Design of an Automated Purification System for Biologically-Active
Macromolecules
by
Eric Hoarau
B.S., Mechanical Engineering
University of California at Berkeley, 1999
Submitted to the Department of Mechanical Engineering
in partial fulfillment of the requirements for the degree of
Master of Science in Mechanical Engineering
at the
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
ENO
June 2001
© Massachusetts Institute of Technology 2001
All rights reserved
MASSACHUSETTS INSTITUTE
OF TECHNOLOGY
JAN 2 3 2002
LIBRARIES
Signature of Author ................................
........................................
Department of Mechanical Engineering
May 11, 2001
C ertified by ...................
......
......................
...................
Kamal Youcef-Toumi
Professor of Mechanical Engineering
Thesis Supervisor
Accepted by ..................................................
Ain A. Sonin
Chairman, Department-Committee on Graduate Students
Design of an Automated Purification System for Biologically-Active
Macromolecules
by
Eric Hoarau
Submitted to the Department of Mechanical Engineering
on May 11, 2001, in partial fulfillment of the
requirements for the degree of
Master of Science in Mechanical Engineering
Abstract
All biologically active macromolecules (BAMs) including pharmaceutical drugs need
purification as part of their production process to ascribe therapeutic properties. Therefore, the
purification of BAMs has proven to be one of the fundamental challenges in drug production and
discovery. There are different types of techniques presently being used to purify BAMs: manual,
mechanical, or a combination of the two. Although purification techniques have improved
dramatically in the last few years, the labor hours, chemicals used, delays and imprecision are
still limiting factors.
This thesis first presents a stain-free ultraviolet absorption detection method that can be used
to detect a variety of biologically active macromolecules in an electrophoresis gel. Next, it
describes the design of a system that will automate the recovery and storage of samples from an
electrophoresis gel.
The following components, which were designed and built for this system, are described in
this thesis. An ultraviolet light source outputs a wide, collimated monochromatic beam of light to
detect specific molecules within the gel. A mechanical cutting device excises bands of various
shapes and sizes from an electrophoresis gel. A transportation system was developed to rapidly
move the cutting device between the different stations. A cleaning station was implemented to
clean and store the cutting tips that are used to excise the gel. Finally, a temperature-controlled
storage station was developed to store the excised samples until needed for further analysis.
Thesis Supervisor: Kamal Youcef-Toumi
Title: Professor of Mechanical Engineering
2
Acknowledgments
Thank you Professor Kamal Youcef-Toumi for supervising this thesis and for your support and
guidance during my time at MIT. I would like to extend my gratitude to Alpine Pharmaceuticals,
Inc. for funding this research. In particular, I would like to thank Dr. Manzoor Shah for his
assistance in completing this work.
I would like to thank Belal, Bernardo, Byron, Namik, and Osamah for their help on this
project. I also thank my colleagues in the Mechatronics Research Laboratory and the Intelligent
Machine Laboratory who have made my time there enjoyable. Thanks to Leslie and Carolyn for
their help with cumbersome MIT paperwork. I would also like to thank Arin, May-Li, and Mike
for proofreading this thesis.
Finally, I would like to thank my family for their continuous encouragement.
3
Contents
Chapter 1
Introduction..............................................................................................................13
1.1
Background Inform ation...............................................................................................
13
1.2
Gel Electrophoresis Principle .......................................................................................
14
1.3
Purpose of the Research...............................................................................................
16
1.4
Functional Requirem ents and Scope of Research........................................................
16
1.4.1
Functional Requirem ents........................................................................................
16
1.4.2
Scope of Research .................................................................................................
17
Chapter 2
D ifferent V isualization Techniques......................................................................
18
2.1
Overview ..........................................................................................................................
18
2.2
Laser Induced Fluorescence (LIF) ..............................................................................
20
2.3
UV Absorption.................................................................................................................22
2.4
Com parison .....................................................................................................................
24
2.5
Proposed Technique.....................................................................................................
25
Chapter 3
Presentation of the M ain C om ponents..................................................................
26
3.1
Overall Overview .............................................................................................................
26
3.2
Detection System ......................................................................................................
27
3.3
Precision X Y stage...........................................................................................................28
3.4
Variable Light Source ...................................................................................................
29
3.4.1
Specifications ............................................................................................................
29
3.4.2
Existing D esigns...................................................................................................
31
3.4.3
Lam p Choice .............................................................................................................
33
3.4.4
Different Approaches to Filtering .........................................................................
38
3.4.5
Different D iffraction Gratings...............................................................................
42
4
3.4.6
Optical Components.............................................................................................
45
3.4.7
Final D esign and Lam p Characteristics .................................................................
55
3.4.8
Recom m endations and Future Work....................................................................
61
3.5
Cutting Tool.....................................................................................................................65
3.5.1
Specifications ........................................................................................................
65
3.5.2
Existing D esigns....................................................................................................
65
3.5.3
Possible D esigns....................................................................................................
66
3.5.4
D esign of the Cutting device..................................................................................
68
3.5.5
Results and Recom mendations.............................................................................
71
D esign of the Cutter Transportation System ................................................................
74
3.6
3.6.1
Requirem ents.............................................................................................................74
3.6.2
Possible D esigns....................................................................................................
74
3.6.3
D esign of the Transportation System ....................................................................
75
3.6.4
Results and Recom mendations.............................................................................
77
Cutting Tips Changing Station......................................................................................
78
3.7
3.7.1
Requirem ents.............................................................................................................78
3.7.2
D esign of the Tool Changing Station....................................................................
78
3.7.3
Results and Recom mendations.............................................................................
80
Cutting Tip Cleaning Station ........................................................................................
82
3.8
3.8.1
Requirem ents.............................................................................................................82
3.8.2
Possible Cleaning M ethods ...................................................................................
82
3.8.3
D esign of the Cleaning Station.............................................................................
84
3.8.4
Results and Recom m endations...............................................................................
85
Temperature Controlled Sam ple Storage Station ........................................................
90
3.9
3.9.1
Requirem ents.............................................................................................................90
3.9.2
Design of the Temperature Controlled Storage Station ........................................
90
3.9.3
Results and Recom mendations...............................................................................
92
Chapter 4
C onclusion ...............................................................................................................
5
97
A ppendix A ...........................................................................................................
101
Al
M achine Pictures ...........................................................................................................
102
A2
Parts List ........................................................................................................................
104
A3
Vendors Inform ation......................................................................................................
108
A4
Circuit Schem atic of Selected Components ..................................................................
110
Appendix B ...........................................................................................................
112
B
CCD Cam era Specifications..........................................................................................
113
B2
XY Stage Specifications................................................................................................
116
Appendix C
The Light Source ...................................................................................................
118
Cl
Assembly Drawings .......................................................................................................
118
C2
Parts Drawings...............................................................................................................
123
C3
Purchased Parts..............................................................................................................
147
Appendix D
The Excision D evice.............................................................................................151
Dl
Assembly Drawings.......................................................................................................151
D2
Parts Drawings...............................................................................................................153
D3
Purchased Parts.................................................................................................
D4
The protean 2-D Spot Cutter by Bio-Rad......................................................................
161
Appendix E
The Excision Device Transportation System .......................................................
162
El
Derivations for the transportation system .......................................................................
.... 160
163
E1.1
System Torque Calculation.....................................................................................
163
E1.2
Calculation of System Resolution and Speed .........................................................
165
E1.3
Calculation of Rack and Pinion Specifications.......................................................
165
E2
Parts Drawings................................................................................................................
6
166
Appendix F
The Cutting Tip Changing Station.........................................................................172
Fl
P art D raw ings..................................................................................................................
172
Appendix G
The Temperature Controlled Storage Station........................................................179
GI
Derivations for the Temperature Controlled Station .....................................................
180
G1.1
Heat Load Calculations ...........................................................................................
180
G1.2
Calculation of Additional Parameters .....................................................................
181
G1.3
System Parameters ..................................................................................................
182
G2
Thermoelectric Cooler Data Sheet.................................................................................
183
G3
Assembly of the Thermoelectric Cooler Storage Station ..............................................
184
Appendix H
The System Software Interface..............................................................................186
Hi
The Interface Screens.....................................................................................................187
7
List of Figures
Figure 1.1: Schematic of the electric field between two parallel plates .................................................
14
Figure 1.2: M acromolecules mobility in matrix gel versus size............................................................
15
Figure 1.3: Example of separation of molecules using electrophoresis.................................................
15
Figure 2.1: Typical LIF detection system [3].............................................................................................
20
Figure 2.2: Instrumental set-up of a capillary electrophoresis system. [2]............................................
21
Figure 2.3: Absorption of light by a sample. [4] ....................................................................................
22
Figure 2.4: CCD based DNA detection in systems for electrophoresis gels. [9]....................................
24
Figure 2.5: Quantum Efficiency of several CCD Arrays............................................................................
25
Figure 3.1: Drawing of the purification system.....................................................................................
26
Figure 3.2: Detection system by: a) taking a picture, b) scanning the gel..................................................
27
Figure 3.3: 402 LN series XY table from Daedal. [4]................................................................................
28
Figure 3.4: Double stranded DNA absorbance spectrum. ......................................................................
29
Figure 3.5: Absorbance and light source spectra.....................................................................................
30
Figure 3.6: Spectroscopic light system..................................................................................................
31
Figure 3.7: 254nm Ultralum transiluminator..............................................................................................
32
Figure 3.8: Proposed light source...............................................................................................................
32
Figure 3.9: Comparison of lamp output stability.....................................................................................
33
Figure 3.10: Spectrum of the L2D2 deuterium lamp using the CCD detector. .....................................
34
Figure 3.11: Deuterium lamp test: a) apparatus, b) and c) test results...................................................
35
Figure 3.12: Lamp intensity at 254nm versus the angle.............................................................................
36
Figure 3.13: Lamp intensity versus the angle for each bulb...................................................................
37
Figure 3.14: Filter wheel approach.............................................................................................................
38
Figure 3.15: Variation of wavelength with incidence angle..................................................................
39
Figure 3.16: Diffraction grating approach .............................................................................................
39
Figure 3.17: Diffraction grating [11]..........................................................................................................
40
Figure 3.18: Grating driving mechanism. [11]......................................................................................
41
Figure 3.19: Overlapping of spectral orders [11] ....................................................................................
41
Figure 3.20: Plane grating mounts. [11]..................................................................................................
42
Figure 3.21: Concave grating mount [11]................................................................................................
43
8
Figure 3.22: Optics set-up to focus and collimate the light ...................................................................
45
Figure 3.23: Light source set-up using a reflector...................................................................................
46
Figure 3.24: Light source set-up with direct grating's illumination........................................................
47
Figure 3.25: Light source illumination at different wavelengths for set-up with lenses. .......................
49
Figure 3.26: Light source illumination at different wavelengths for set-up with reflector......................
50
Figure 3.27: Light source illumination at different wavelengths for set-up with direct illumination......... 51
Figure 3.28: Light source spectrum analysis for three different focusing set-ups.................
52
Figure 3.29: Proposed light source .............................................................................................................
55
Figure 3.30: Set-up to measure unwanted reflected light. ......................................................................
58
Figure 3.31: Influence of bare aluminum walls on the output signal. ....................................................
58
Figure 3.32: Output power of the variable light source..........................................................................
59
Figure 3.33: Illumination profile of light source at 260nm. ....................................................................
60
Figure 3.34: Peak location throughout the beam .....................................................................................
60
Figure 3.35: Possible Gaussian addition to explain the spectrum shape ....................................................
61
Figure 3.36: Deuterium lamp light path .....................................................................................................
61
Figure 3.37: Direct illumination with a baffle to restrict the light reaching the grating.........................
62
Figure 3 38: Results from testing the influence of the baffles on the direct illumination set-up....... 63
Figure 3.39: Projecting type deuterium lamp. ............................................................................................
64
Figure 3.40: Potential design of the cutting device using a:..................................................................
67
Figure 3.41: The cutting device..................................................................................................................
69
Figure 3.42: The cutting tip ........................................................................................................................
71
Figure 3.43: Cutting tips cross section ..................................................................................................
72
Figure 3.44: Band expulsion test result ..................................................................................................
72
Figure 3.45: Possible modifications of the cutting tip expulsion system ..............................................
73
Figure 3.46: Transportation systems: a) pulley-belt, b) lead screw, c) rack-pinion ...............................
74
Figure 3.47: The cutter transportation system.......................................................................................
75
Figure 3.48: The cutting tip storage station...........................................................................................
79
Figure 3.49: Tool changing station solenoid input voltage versus time .................................................
81
Figure 3.50: Other designs to increase number of stored cutting tips .....................................................
81
Figure 3.51: Different cutting tip cleaning methods..............................................................................
83
Figure 3.52: The UC-1 ultrasonic cleaner .............................................................................................
85
Figure 3.53: Ultrasonic cleaner assembly using one beaker for cleaning and one for rinsing ...............
89
Figure 3.54: Schematic of a thermoelectric cooler ................................................................................
90
Figure 3.55: The temperature-controlled sample storage station............................................................
91
9
Figure 3.56: Temperature on the thermoelectric cold side during power-up and power-down...............
92
Figure 3.57: Temperature at a height of 10-mm with different heat transfer materials............. 93
Figure 3.58: Cooling test, temperature at a height of 10-mm with an aluminum block.............
95
Figure 3.59: Heating test, temperature at a height of 10-mm with an aluminum block .............
95
Figure Al: The designed automated purification system for biologically active macromolecules.......... 102
Figure A2: Pictures of selected components.............................................................................................
103
Figure A3: Schematic for the 4SQ-120BA34S stepper motor .................................................................
110
Figure A4: Schematic for the L92121-P2 stepper motor..........................................................................
111
Figure A5: Schematic for the solenoid....................................................................................................
111
Figure A6: Schematic for one of the feedback sensor ..............................................................................
111
Figure B 1: 402 LN Series XY Table from Daedal. [4] ............................................................................
116
Figure Cl: Assembly of the light source outside walls ............................................................................
119
Figure C2: Assembly of the light source components #1.........................................................................
120
Figure C3: Assembly of the light source components #2.........................................................................
121
Figure C4: Assembly of the diffraction grating mechanism.....................................................................
122
Figure C5: Drawing of the light source cover plate..................................................................................
124
Figure C6: Drawing of the light source front wall.................................................................................
125
Figure C7: Drawing of the light source sidewall......................................................................................
126
Figure C8: Drawing of the light source back wall....................................................................................
127
Figure C9: Drawing of the light source base plate #1 ..............................................................................
128
Figure C10: Drawing of the light source base plate #2............................................................................
129
Figure C11: Drawing of the light source wall blocks...............................................................................
130
Figure C12: Drawing of the light source inside wall #1...........................................................................
131
Figure C13: Drawing of the light source inside wall #2...........................................................................
132
Figure C14: Drawing of the light source bracket for the inside wall........................................................
133
Figure C 15: Drawing of the light source shutter holder ...........................................................................
134
Figure C16: Drawing of the light source heat sink...................................................................................
135
Figure C17: Drawing of the light source base plate for the deuterium lamp............................................
136
Figure C18: Drawing of the light source base for the diffraction grating mechanism .............................
137
Figure C19: Drawing of the light source diffraction grating holder.........................................................
138
Figure C20: Drawing of the light source diffraction grating shaft ...........................................................
139
Figure C21: Drawing of the light source diffraction grating bearings spacer ..........................................
140
Figure C22: Drawing of the light source diffraction grating shaft arm....................................................
141
Figure C23: Drawing of the light source diffraction grating base leg #1.................................................
142
10
Figure C24: Drawing of the light source diffraction grating base leg #2.................................................
143
Figure C25: Drawing of the light source diffraction grating motor bracket.............................................
144
Figure C26: Drawing of the light source diffraction grating motor holder ..............................................
145
Figure C27: Drawing of the light source diffraction grating motor coupling ..........................................
146
Figure Dl: Assembly of the excision device............................................................................................
152
Figure D2: Drawing of the main frame ....................................................................................................
154
Figure D3: Drawing of the small frame....................................................................................................
155
Figure D4: Drawing of the cutting tip holder...........................................................................................
156
Figure D5: Drawing of the cutting tip base ..............................................................................................
157
Figure D6: Drawing of the stopper block.................................................................................................
158
Figure D7: Drawing of the flexible shaft plate.........................................................................................
159
Figure E1: Schematic of the rack and pinion system ...............................................................................
163
Figure E2: Torque curves for the 4SQ-120BA34S stepper motor ..........................................................
164
Figure E3: Assembly of the transportation system ...................................................................................
167
Figure E4: Drawing of the transportation system main frame .................................................................
168
Figure E5: Drawing of the transportation system carriage frame #1........................................................
169
Figure E6: Drawing of the transportation system carriage frame #2........................................................
170
Figure E7: Drawing of the transportation system rack.............................................................................
171
Figure Fl: Assembly of the cutting tip changing station..........................................................................
173
Figure F2: Drawing of the base plate .......................................................................................................
174
Figure F3: Drawing of the end-piece........................................................................................................
175
Figure F4: Drawing of the shaft holder ....................................................................................................
176
Figure F5: Drawing of the sliding arm .....................................................................................................
177
Figure F6: Drawing of the shaft ...............................................................................................................
178
Figure G 1: Diagram of the heat transfer in the storage system ............................................................
180
Figure G2: The temperature controlled storage station............................................................................
184
Figure G3: Construction of storage tank: a) frame only, b) with the insulation.......................................
184
Figure G4: Assembly of the thermoelectric base: a) top view, b) bottom view .......................................
185
Figure HI: Sample of the interface control for the cutting device, transportation system, and light source
...........................................................--..............................................................................................
Figure H2: Sample of the interface control for the XY stage..................................................................
11
18 7
187
List of Table
Table 1.1: Comparison table for the different visualization techniques [2]..............................
19
Table 3.1: Different available UV lamps. ..................................................................................
33
Table 3.2: Selected concave holographic grating from Richardson grating laboratory............44
Table 3.3: Results of the spectrum bandwidth and location test...............................................54
Table 3.4: Different surface reflectivity...................................................................................
58
Table 3.5: Results of the ultrasonic test for different solution.................................................
86
Table 3.6: Results of the ultrasonic test for different solution.................................................
87
Table 3.7: Test result for blade cleaned while touching the bottom of the tank .......................
88
Table 3.8: Selected material property at 21'C..........................................................................
94
12
Chapter 1
Introduction
1.1 Background Information
The development of new pharmaceuticals requires the use of pure biological samples, which
allow for a thorough analysis of the therapeutic properties of newly developed drugs. Different
methods are used to purify these samples such as chromatography, gel electrophoresis, capillary
electrophoresis (CE), and high performance liquid chromatography (HPLC).
Each method has its advantages and disadvantages.
Chromatography techniques are time-
consuming, typically requiring five to seven working days, and are also imprecise, thus prone to
errors. Although they are routine procedure, gel electrophoresis methods also require intensive
labor -at least two or three working days and they result in a 30-40% product loss. The recovery
of the sample is also problematic and imprecise. Currently, both these processes are carried out
manually, which creates some repeatability and contamination issues.
This thesis develops the design of a system that will automate the recovery and storage of
biological samples in an electrophoresis gel. It will also address the issues of increasing the
efficiency and repeatability of the electrophoresis process, and eliminate possible sample
contamination.
This first chapter provides a background and explanation of the electrophoresis process. It
also states, in further detail, the objectives of the robotic system.
Chapter 2 describes and compares different possible visualization techniques, and presents
the selected detection method.
Chapter 3 presents the design of each component, and also shows the performance and
possible improvements for each.
13
1.2 Gel Electrophoresis Principle
Electrophoresis is a method used for the separation of biological macromolecules (such as
proteins, nucleic acids, carbohydrates, and lipids) based on their size, charge, and conformation.
Most biological molecules have an electrical charge, which is dependent on the molecule itself
and on the pH of the solution surrounding it. A molecule that is placed in an electric field (see
Figure 1) moves toward the cathode if negatively charged or toward the anode if positively
charged. Molecules of different charges migrate at different rates.
Figure 1.1: Schematic of the electric field between two parallel plates
If the distance between the plates is d and the potential difference is E, then the force, F,
exerted on a molecule of charge, q, moving in an electric field is
F =(
d
q
(1.1)
Since the molecule of radius, r is moving in a medium of viscosity p, this force is equal to
the frictional force, Ff, (assuming that the molecule is moving at a constant velocity, V).
Ff = 6zrpV = F
(1.2)
V=
(1.3)
Eq
61rad
Most electrophoresis techniques use a gel (polyacrylamide or agarose), a paper, or cellulose
acetate as the supporting media. The matrix of the gel creates a sieving effect that interferes with
the molecule's motion and therefore increases the separation gap between molecules of different
sizes. As a result, larger molecules migrate slower. The velocity equation above does not
account for the sieving effect due to the pores in the gel. It is more accurate to use the mobility
of the molecules to describe their motion. The mobility of the molecule in a gel matrix exhibits
the characteristic profile shown in Figure 1.2. The gel concentration should be adjusted so that
the molecule mobility lies in the linear region of the plot. This will ensure a more uniform
spread of the molecule bands along the gel lane as seen in Figure 1.3. [1]
14
Log molecular size
Figure 1.2: Macromolecules mobility in matrix gel versus size
Anode
Anode
A
Electrophoresis
A
Large
gel
[
Medium
0 Small
o
0
A
An
A
AA
o
0
o 0
o
0 0
Cathode
Cathode
After electrophoresis
Before electrophoresis
Figure 1.3: Example of separation of molecules using electrophoresis
Once separated, the molecules are invisible to the naked eye. A staining process is usually
done to detect them in the gel. Several staining methods are available. Molecules can be stained
with an intercalating dye such as ethidium bromide.
This stain is commonly used but is
hazardous. Silver stains may also be used. They are the most sensitive but molecules cannot be
recovered afterwards. The gel can also be blotted onto a nitrocellulose filter by electrophoresis
and visualized using an x-ray film or other staining procedures.
This method requires
radioactive labels and is therefore highly hazardous. Another method used does not require any
stains, and is know as UV shadowing. However, it requires a larger concentration of the sample
than other methods, and is applicable specifically to DNA molecules. [1]
15
1.3 Purpose of the Research
As stated previously, current gel electrophoresis methods have several drawbacks.
stains need to be used to detect the molecules in the gel.
Namely,
Most intercalating stains bond
physically to the molecule, so de-staining is necessary if further analysis of the molecule is
desired. De-staining is a long, inefficient, costly and hazardous process. Afterwards, the desired
bands need to be excised from the gel. Since this is done manually by an operator, it creates
problems. The operator is exposed to hazardous chemicals and must exercise a large amount of
caution in his work. Also, due to low precision, the operation is not repeatable, and the sample
might be lost. Furthermore, during the process, an unclean blade or a handling mistake could
contaminate the sample.
In summary, six problems exist in current electrophoresis methods:
1
De-staining of desired bio-molecules is a costly and difficult process.
2
Electrophoresis is a slow process.
3
Large product losses occur during the operation.
4
Health concerns arise since the operator handles hazardous chemicals.
5
Procedure is prone to human error.
6
Method has low repeatability.
1.4 Functional Requirements and Scope of Research
The aim of this project is to design a robotic purification system for biologically active
macromolecules. This machine is intended to combine the efficiency of a gel electrophoresis
system with the precision of a mini-robot, and thus automate the whole electrophoresis process.
1.4.1
Functional Requirements
From a functional standpoint, this machine will have four-fold applications:
First, it will automate the manual electrophoretic procedures, which are routinely used all over
the world by numerous scientific personnel. Specifically, the mini-robot system will speed up the
purification of Biologically Active Macromolecules (BAM) by:
a) reducing the number of steps and automating repetitive steps,
b) increasing the amount of end product by reducing the waste of the BAM,
c) eliminating contact contamination.
16
Second, it should have the capability to:
a) determine molecular weights of BAMs,
b) quantify concentrations of BAMs,
c) record/store data for photo-imaging.
Third, the built-in artificial intelligence of the machine should enable it to adjust to various
situations. It should also allow a scientist to operate an experiment from a remote position,
access data in real-time, and change experimental parameters at his/her will.
Fourth, it should have the capability to provide 24-hour, real-time, on-line access to a repository
of molecular data (protein/DNA sequences, structural homologues, physico-chemical properties
etc.) once an experimental run is complete.
1.4.2
Scope of Research
This research is composed of two main parts. The first part addresses the development of a stain
free detection system that can be used for a variety of BAMs. The second part involves of the
design of the main components of the robotic system, namely:
*
Variable ultraviolet light source
"
Mechanical cutting device to excise desired bands
"
Transportation system for the cutting device
* Washing station to clean and store cutting-tip ends
"
Temperature controlled sample storage station
17
Chapter 2
Different Visualization Techniques
2.1 Overview
This chapter introduces the different techniques to visualize bio-molecules in a gel. The two
most applicable to our set-up are reviewed, then after comparing them, the selection is made.
There are several detection methods for gel electrophoresis: absorbance, amperometry,
conductometry, fluorescence, and indirect methods. The selection of an appropriate method is
dependent on the application. The specifications for the detection system are:
*
No stains should be used, so that further analysis of the sample is possible. This will also
increase the efficiency of the process.
*
The method should be applicable to a variety of BAMs: nucleic acids, proteins, and
carbohydrates.
*
The visualization of the bands in the gel should be fast enough to avoid photo bleaching
and gel shrinkage. Photo bleaching is a change in the chemical structure of a molecule
due to an intense or prolonged ultraviolet light exposure. Gel shrinkage occurs as the gel
dries. This is caused by either by gel dehydration or by an increase in temperature due to
UV light exposure. Both these phenomena should be avoided since they cause a change
in the shape of the gel, and thus result in an inaccurate band location.
*
The method should have enough resolution to be able to detect sample amounts less than
500 nanograms.
18
Detection method
UV Absorbance
Applications
Characteristics
Nucleic acids,
proteins, peptides,
small ions
Only electroactive
compounds in
complex matrices
Ion analysis
Amperometry
Conductometry
Fluorescence (LIF)
Indirect absorbance
Indirect fluorescence
Equipment needed
Universal and easy to
use but relatively low
sensitivity
High sensitivity and
selectivity, difficult to
establish
Low sensitivity, not
universal
High sensitivity,
selective, expensive,
not universal
Universal, low
sensitivity, restriction
of buffer choice
Universal, high
sensitivity, restriction
Amino acids, nucleic
acids, peptides,
proteins
Ion analysis,
carbohydrates
Only non-fluorescent
compounds
of buffer
Variable UV lamp,
light sensor
Platinum electrodes,
power supply
Conductivity cell,
conductometer
Several lasers,
Photomultiplier tube,
spectroscopy method
Stable UV lamp, light
sensor, dye screen
Stable arc lamp, laser,
filter
I
_I
Table 1.1: Comparison table for the different visualization techniques [2]
A comparison table of the different methods is shown in Table 1.1.
Amperometry and
conductometry methods are usually applied in conjunction with capillary electrophoresis
techniques for the detection of specific compounds that other methods cannot detect.
The
indirect detection methods are primarily used for detecting substances with low absorptivity, that
cannot be detected by their direct detection counterparts.
They are restricted to specific
substances and require the use of a low concentration background electrolyte to work well. They
also need a highly stable light source to ensure a uniform background signal [2]. The last two
methods (absorbance and LIF) satisfy most of the specifications. They could both be applied to
our system and will now be presented. However, it is to be noted that this is only an overview of
the methods. An in-depth analysis is outside the scope of this thesis, and one should consult
appropriate literature for more information.
19
2.2 Laser Induced Fluorescence (LIF)
Laser induced fluorescence is one of the most sensitive detection methods of bio-molecules. It
consists of exposing a sample to electromagnetic radiation (usually with a laser). The molecules
in the gel become excited to higher energy levels, and fluoresce at different wavelengths as they
decay to lower levels.
The higher sensitivity is due to the low background noise of the
fluorescent signal.
Capillary
Focusing
lens
~
Cuvette
Beam block
Laser source
Collection lens
and filter
PMTt
Figure 2.1: Typical LIF detection system [3]
A typical set-up is shown in Figure 2.1. The laser is focused on the sample. A PhotoMultiplier Tube (PMT) senses the fluorescence emission collected by the lenses. A high power
laser is necessary to obtain high resolution. This is because the fluorescence and background
signal increase linearly with the laser intensity.
However, the background noise increases
proportionally to the square root of the laser intensity. Hence, a higher resolution is achieved by
increasing the intensity. Since the laser used as the exciting beam has a small aperture, the LIF
method is primarily used with capillary electrophoresis.
20
acquisition
capilarydata
+
detector
buffer
reservoir
sample
vial
buffer
reservoir
high voltage
power supply
Figure 2.2: Instrumental set-up of a capillary electrophoresis system. [2]
A Capillary Electrophoresis (CE) set-up is shown in Figure 2.2. A capillary tube is filled
with gel and buffer. Then, using a high voltage power supply (up to 30 kV), an electric field is
applied between the two buffer reservoirs. The sample is inserted into the capillary by replacing
one of the buffers with the sample vial. In the detection section of the capillary, the capillary is
usually replaced with a rectangular cuvette. This is necessary, because if the laser were passed
through a round capillary, scattering and image distortion would occur. This would result in
larger background noise and therefore a much lower sensitivity of the system.
The LIF technique allows for great sensitivity, but only at the cost of being expensive and not
universal. Its strength lies in its ability to analyze the electronic structure of the bio-molecules,
and to determine the analyte's concentration, which is proportional to the area under the output
signal curve. Also, LIF can only analyze one sample at a time, and since the capillary has an
internal diameter on the order of 100-micrometers (equal to the diameter of a human hair), this
results in a very low throughput. Systems that can process several capillaries at a time are being
developed, but they are complex and expensive.
21
2.3 UV Absorption
UV absorption is one of the most popular detection methods in use today, even though it has a
lower sensitivity than other techniques. This is due to the fact that it can be applied to a wide
range of bio-molecules, and is easy to use. It consists of exposing a sample to electromagnetic
radiation (usually with a UV lamp). At the location of the sample, the intensity of the UV signal
drops because the molecules absorb UV light. This variation in signal can be detected. The
molecule can also be identified because each bio-molecule has a characteristic absorption
spectrum. The absorption peaks of different molecules are located at different wavelengths. The
absorption peak is around 214nm and 280nm for proteins depending on their type, 230nm for
carbohydrates and peptides, and 260nm for nucleic acids.
The Beer-Lambert Law describes the light absorption principle for a non-opaque sample.
A = exbxc
(2.1)
where
A
b
c
E
is the measured absorbance,
is the path length [cm],
is the analyte concentration [M],
is the wavelength-dependent molar absorptivity coefficient [M- cm-'].
Absorbing sample
D*
of concentration c
Path length b
Figure 2.3: Absorption of light by a sample. [4]
Figure 2.3 presents a schematic of the absorption principle.
The transmittance, T, of a
sample is the quantity usually measured by instruments.
T =Io
10
(2.2)
where
I
10
is the light intensity after absorption,
is the initial light intensity.
22
The relation between the absorbance and the transmittance is:
A = -logT = -log 10
(2.3)
Typical absorption techniques use stains to increase the signal to noise ratio, and photo paper
to record the band location. This is hazardous, costly, and time consuming. There is another set
of absorption methods that does not use stains. They fall under the umbrella of UV shadowing
techniques. The basic method for DNA molecules was described almost 50 years ago [5]. It
consists of placing the gel above a 254nm light source.
Atop the gel, a transparent UV-
fluorescent material such as a standard, 1mm thick, minigel glass plate is placed. The plate will
fluoresce everywhere except where the DNA has absorbed the UV light. The bands appear as
dark regions on a light background. Quantities of unstained DNA in a gel of as low as 0.25
gg/bands have been detected in recent experiments [6]. In another application of UV shadowing,
even smaller quantities were detected. The process involves transferring unstained nucleic acid
to a nylon membrane and then visualizing the bands under UV light. The nylon membrane has a
small UV induced fluorescence. Sensitivity down to 10ng has been achieved [7].
All the methods presented above record the band location on special photographic paper.
The paper is then scanned to obtain an electronic version.
therefore it is not desirable.
This process is time-consuming and
Two other methods have been previously developed that can
visualize the gel directly. They have been developed for visualizing DNA molecules only and
are not applicable to other bio-molecules. The first one is a variation of UV shadowing. It uses a
phosphor storage screen (that fluoresces under UV light) to record the location of the bands in
the gel. The lowest sensitivity obtained with this first method to date is around 400ng [8]. The
second method uses direct absorption to detect the migration of DNA molecules through a gel.
As seen in Figure 2.4, UV light is shone through a gel using a set of fiber optics. On the other
side, a set of fiber optics collects the light and brings it to a Charged Coupled Device (CCD).
Since the CCD camera used is not sensitive in the UV region, it needs to be coated with a
phosphor lumogen coating that absorbs UV light (specifically at 260nm) and fluoresces in the
visible range. The CCD then has a Quantum Efficiency (QE) of 12% at 260nm. With this
system, the lowest sensitivity achieved was 1.25ng [9]. This method is far more sensitive than
the previous one, but it only scans the gel, therefore it has a smaller throughput, even though it
uses multiple fiber optics.
23
electrophoresis
UV
fibre guide
15cm
D2 hmg
Ir_
CCD
1
I
I
I
read-out
I
X~i~K)
1~~~~ 1V4
3 planes of nine I mm illumination fibres
260mu filter
3 planes ofnine 1 mm collection fibres
Figure 2.4: CCD based DNA detection in systems for electrophoresis gels. [9]
2.4 Comparison
After describing laser-induced fluorescence and UV absorption, the following conclusion can be
made. The LIF method has better resolution capabilities than absorption, however it needs stains
to achieve the high sensitivity. On the other hand, UV absorption can be used without stains, but
has lower sensitivity. It can have a higher sensitivity, as explained in the CCD method, but only
at the cost of losing its universality. Since the LIF technique is only applied in conjunction with
capillary electrophoresis, it can sense one or a few samples at once, whereas the absorption
technique allows for a snapshot picture of the entire gel at once.
For our purpose, UV absorption arises as the better solution. However, a new approach to
UV absorption will be taken to increase its efficiency, without losing its universality.
24
2.5 Proposed Technique
Of all the UV absorption methods mentioned previously that use unstained samples, none can be
universally applied to several bio-molecules.
quantum efficiency in the UV region.
This is due to the lack of a detector with high
However, a new technology, known as back-thinned
CCD, has been developed, and it allows for a dramatically improved UV response of the
detector. Figure 2.5 shows the quantum efficiency of different CCD cameras. The back-thinned
CCD has a minimum QE of 45% between 200 and 400 with a maximum of 83 % around 230nm.
Front-sided (UV coated) cameras only have efficiencies of around 8% in the UV region.
40
31:40
0
00
00
10
WAVELENGi1H ftmlI
Figure 2.5: Quantum Efficiency of several CCD Arrays.
(Courtesy of Hamamatsu Inc.)
Using this CCD camera, the gel is visualized directly, and the band location is found by
direct UV absorption measurement. A variable light source (200-400nm with 15nm bandwidth)
was developed to allow for a precise selection of the target molecules to be visualized. This
system can take pictures of a gel at any wavelength in the UV region, and has the potential to
have a higher sensitivity than previous ones. The scanning approach is also investigated since it
can produce information about the structure of the sample.
25
Chapter 3
Presentation of the Main Components
3.1 Overall Overview
In this chapter, the main components of the automated purification system are presented. Figure
3.1 shows the main components of the machine. These are the detection system (CCD camera
and spectrometer), the precision XY stage, the variable light source, the cutting device, the
sample transportation system, and the temperature controlled storage station.
CCD Camera
Chemical Storage
Gel Cutting
Device
XY Stage
I
Figure 3.1: Drawing of the purification system.
26
The operation of the system is as follows. The gel is placed in a tray on top of the UV light
source, which is attached to the XY stage. Once the CCD camera visualizes the gel and the
desired bands to be cut are identified, the stage moves to place the first band to be excised under
the cutting tool. The cutter excises the band and carries it to the temperature controlled storage
area. The sample is expelled into a storage vial using compressed air. Then, the cutting tool
moves to the cleaning station to exchange the used cutting tip with a clean one. Next, the cutter
tool performs the next band excision, while the contaminated cutting tip is cleaned using an
ultrasonic cleaner. This cycle is repeated until all the desired bands have been extracted from the
gel.
Now that the general operation of the machine has been explained, each individual
component will be presented.
3.2 Detection System
The detection system was explained in the previous chapter. There are two different set-ups used
in our machine. The first one takes snapshot pictures of the gel to locate the band shape and
position inside the gel, while the second one scans the gel to obtain the absorption spectrum of a
specific band. The two systems are shown in Figure 3.2.
CCD camera
Spectrometer
CCD camera
Fiber
optic
Uv lens
Computer
Light source
Computer
a)7
Light source
Gel
b)E_1
Figure 3.2: Detection system by: a) taking a picture, b) scanning the gel.
The CCD camera, spectrometer, fiber optic, and controlling software were purchased from
Acton research. The light source was developed and is presented in a section 3.4. The CCD
camera uses a back-illuminated and UV-coated Hamamatsu CCD with a 1024 x 256 pixel
format. As mentioned in the previous chapter, this CCD camera was chosen because it has a
high quantum efficiency (43%-85%) in the UV region (200-400nm). The system comes standard
with a 100-kHz, 16-bit analog-to-digital (ADC) converter and a 12-bit, 1-MIHz ADC for rapid
kinetics and fast system alignment. The spectrometer is an Acton Research SpectraProl50. Two
27
1200 1/mm gratings are included, a 300nm blazed for the UV range and a 500nm blazed for the
visible range. The software used to control the CCD camera and the spectrometer is Princeton
Instrument WinView for image acquisition and WinSpec for scanning.
To visualize the gel
using the CCD camera, a model UV8040B lens (78mm, F/3.8, UV imaging lens) from Universe
Kogaku Inc is used. The technical sheets of the CCD detector and the UV lens are shown in
Appendix B 1.
3.3 Precision XY stage
The XY stage is used for both placement of the gel under the CCD camera and for a precise
placement of the band to be excised under the cutting tool. The stage is a model 4020006 XY
table from Daedal. See Figure 3.3. It is designed for repeatable precision positioning of light
payloads over short travels, and can be utilized in applications requiring horizontal, inverted, or
vertical translation. The stage has 150-mm travel in both X and Y directions. It has a step size of
.1 gm, a positional accuracy of 75 gm and a positional repeatability of 12 gm. Since the smallest
expected width of the band to be excised is around 1 mm, this accuracy of .075mm is sufficient
to achieve the desired positional accuracy. The technical specification and the drawings of the
stage are presented in Appendix B2.
The stepper motors to drive the stages are from Compumotor. A digital 1/0 card is used to
generate and send pulses to the power amplifier, which is connected to the stepper motors.
Figure 3.3: 402 LN series XY table from Daedal. [4]
28
3.4 Variable Light Source
As explained previously, each bio-molecule has a different absorption spectrum, and thus a
different absorption peak. Since a direct UV absorption method is used, there is a need for a
compact variable light source capable of focusing on specific wavelengths.
3.4.1
Specifications
The specifications for the light source are summarized first, and then are explained in more
detail. They are:
*
Variable over 200-400nm range.
*
5nm-l5nm bandwidth.
*
20 x 40mm light beam area (similar to detector's shape).
*
Uniform spatially and temporally.
*
Compact to fit on the XY stage.
Most of the bio-molecules of interest have their absorption peak in the UV region of the
spectrum. Specifically, the peak is around 214nm and 280nm for proteins depending on their
type, 230nm for carbohydrates and peptides, and 260nm for nucleic acids. The absorption
spectrum might also be needed to fully characterize a molecule; therefore a range of 200-400nm
is desired for the light source. The absorption spectrum of a double stranded DNA sample is
shown in Figure 3.4. The molecule absorbs strongly at 260nm and below 215nm. The lower
wavelengths are not often used to measure absorption because the agarose gel and the buffer
absorb also below 230nm.
58iI
~Date:
-6p
0T E OF NIWCUE
Z4U1/9.
HOIEK
Tim.: 16:49
PBLUESCRIPT
C AbS]I
28G.8
Wavelength (no)
358.8
Figure 3.4: Double stranded DNA absorbance spectrum.
(Courtesy of http://www.cbs.dtu.dk/dave/roanoke/genetics980211.html)
29
The bandwidth is defined as the width of the peak at half its height. See Figure 3.5. The
bandwidth of the light source is dependent on its application.
For locating the band using
absorption measurement, a bandwidth of half or less than half of the molecule absorption
bandwidth is necessary. However, if the absorption spectrum is desired, then the bandwidth of
the light source needs to be set to one-tenth the absorption bandwidth of the molecule. This is to
ensure that no spectral details are lost. The reason for the larger bandwidth when locating the
band is that the medium surrounding the molecules (buffer or gel) can cause a shift in the
molecule absorption peak anywhere from 1-5nm. Therefore, a larger bandwidth will ensure that
the band is detected.
Another factor to take into account is the signal to noise ratio.
The
intensity of the signal increases as the bandwidth increases. Since the noise level is constant in
the system, the signal to noise ratio increases too. Consequently, it is necessary to have the
largest possible bandwidth that is within the specification to increase the imaging system
resolution. The bandwidth of some of the bio-molecules of interest is about 30-45nm for nucleic
acids, and 25nm for proteins. Therefore, our system should have a bandwidth of around 15nm
for the imaging part and around 3-5nm for the absorption spectrum analysis.
AMolecule
bandwidth
Absorbance
peak
Light
bandwidth
h/ peak
height
200
250
wavelength (nm)
300
Figure 3.5: Absorbance and light source spectra.
The light source output needs to be stable over time in order for the measurement to be
repeatable.
This also allows for good time based measurement, and for baseline correction
methods to be used when visualizing the gel. Moreover, the noise level should be as small as
possible to ensure a good resolution. Finally, the light source needs to be uniform spatially.
Spatial uniformity means that the intensity is uniform across the beam, and that the spectrum
peak shape is be the same throughout the beam area. Errors in the latter point are introduced by
aberrations in the optical system, which will be discussed in Section 3.4.6.
30
3.4.2
Existing Designs
There are a variety of ultraviolet light sources available, however none fit our specifications.
They can be categorized into two groups; spectroscopic and transilluminator. The spectroscopic
systems are made of standard modular components - a light source, optics, and a
monochromator- as seen in Figure 3.6.
Power supply
Monochromator
Lamp
Optics
Motor drive
Fiber optic
Figure 3.6: Spectroscopic light system.
The spectroscopic systems allow for a chromatic tuning of the light, however they are bulky
and expensive. More importantly, they have a small exit aperture (0.1 to 1mm) since they are
primarily used to focus the light into a fiber optic bundle. They can have bandwidths as small as
lnm, which results in low output power. Their purpose is to direct a high intensity UV-light
onto a small sample for spectroscopic analysis.
Transilluminators are used for imaging gels as shown in Figure 3.7. They have a large output
area and are compact (one unit), however they have a fixed wavelength light output.
Furthermore, the illumination is not uniform since the transilluminator uses a series of parallel
tube lamps, and its stability is marginal. Figure 3.7 shows a transilluminator from Ultralum that
has a 254nm light output. This type of light source is typically used for illuminating a gel to
visually locate its bands. Fixed wavelength transilluminators usually cost 4-5 times less than the
least expensive spectroscopic system.
31
Figure 3.7: 254nm Ultralum transiluminator.
(Courtesy of Ultralum, Inc)
These UV light systems cannot be used in our application since they do not fulfill some
important specifications.
The spectroscopic system is large and has a small output beam,
whereas the transilluminator has a fixed wavelength and a non-uniform output. Therefore, the
design of a custom variable light source that meets the aforementioned specification is needed.
Figure 3.8 shows a drawing of the proposed light source.
Electronic
Shutter
Slit
Concave Grating +
Motorize(d Assembly
Collimating
Optics
Deuterium
Lamp
Figure 3.8: Proposed light source.
The variable light source is composed of three main components: the lamp, the filtering
system (concave grating and motorized system), and the optics to focus and collimate the light.
32
3.4.3
Lamp Choice
A lamp that can provide a stable continuous output in the UV range is needed for the light
system. There are four types of UV lamps available: deuterium, xenon, xenon-mercury, and
hollow cathode. Table 3.1 shows a summary of their properties. The deuterium lamp is the ideal
choice for the desired application since it provides a highly stable continuous ultraviolet output
with little visible and infrared emission.
Lamp Type
Wavelength
Spectrum
Deuterium (L2D2)
Xenon
Xenon-mercury
Hollow cathode
185 - 400
185 - 2000
185 - 2000
193 - 852
Stability
Price
(% p-p)
(nm)
Continuous broad
Continuous broad
Continuous broad
Line
0.05
1
2
N/A
Cheapest
Expensive
Expensive
N/A
Table 3.1: Different available UV lamps.
(Courtesy of Hamamatsu Inc)
A L2D2 deuterium lamp and a C4545 power supply from Hamamatsu are used. L2D2 lamps
have a lifetime twice as long as conventional deuterium lamps, around 2000 hours. Moreover
they are 1.3 times brighter and have a higher stability and lower drift as seen in Figure 3.9.
Fluctuation:0.05%p-p
CONVENTIONAL
TYPEr
TIME 10SadIV.)
Figure 3.9: Comparison of lamp output stability
(Courtesy of Hamamatsu Inc)
There are a variety of lamp sizes and shapes. Types L6311-50 (0.5mm aperture) and L631250 (1mm aperture) are selected because they have the smallest size and are already mounted to a
base. This facilitates mounting and more importantly alignment. The 0.5mm aperture has a high
brightness arc, whereas the 1mm aperture provides a more uniform distribution. The selection
between the two types is made through experiments.
To obtain the performances highlighted above, the lamp power supply needs to provide a
constant current for the main power supply section and a constant voltage for the filament power
supply. The Hamamatsu C4545 power supply is specifically designed for these lamps, and
33
therefore is used in the system. Technical information for these components is presented in
Appendix C3.
To select the best lamp for our system, each was characterized by looking at its actual
spectrum and its light distribution. Figure 3.10 shows the spectrum of the L6312 lamp taken
using the spectroscopy system. L6311 spectrum is the same as the L6312 except for a slightly
larger amplitude, therefore it is not shown.
Bulb L6312-50 (1 mm aperture), Intensity at 100mm
10'
3
2.5 -
---
---
1
0.5
-
200
-
250
30
350
400
450
Wavelength (nm)
500
550
600
650
Figure 3.10: Spectrum of the L2D2 deuterium lamp using the CCD detector.
The spectrum shown in Figure 3.10 was obtained at a distance of 100mm directly in front of
the lamp. It exhibits some variation from the spectrum advertised by Hamamatsu. The curve
from 210-400nm should have a smooth decrease. The spectrum taken with the CCD camera in
Figure 3.10 shows bumps around 240nm, 300nm, and 370nm. The CCD detector causes these
bumps. The CCD quantum efficiency is wavelength dependent. As seen in Figure 2.5, the QE
curve has sharp variations in the UV region. The bump locations correlate perfectly with the
sharp changes in the QE of the CCD.
One should be aware that each optical element that
interacts with the light changes the shape of the spectrum because of its wavelength dependent
quantum efficiency. Therefore, these components should be chosen to ensure a uniform intensity
distribution in the wavelength range of interest (200-400nm in the proposed design).
34
Distance
1000
100
50
0,00
a)
Deuterium lamp
-10*
LightFxdfbe
output
optic detector
is rotated around
its center axis
Spectrometer
Bulb 6312-50 (1 mm aperture) Intensity at Angle (-15 to 0 degrees)
x 10"
3
angles -12.5 to 0 degrees
are close to each others
100
-
1.5
Increasing
1
(-15 to 0 degrees)
Step sire: 2.5 degree
0.5
200
250
300
350
400
450
500
550
600
650
Wavelength (nm)
x
10 4
Bulb 6312-50 (1 mm aperture) Intensity at Angle (0 to 15 degrees)
3 1
2.5
Increasing
(0 tQ 15 degrees)
Step size: 2.5 degrees
2
S1 .5
-
-
-- -
--
- -- --
-
-
-
- -
-
C)
01
200
250
300
350
400
450
Wavelength (nm)
500
550
600
Figure 3.11: Deuterium lamp test: a) apparatus, b) and c) test results
35
650
Next, the light distribution of the bulb was determined by measuring the intensity variation at
different angles relative to the lamp center as shown in Figure 3.1 la. The spectrum of the lamp
was recorded every 2.5 degrees from -15 to 15 degrees and analyzed using Matlab.
From the results shown in Figure 3.11 (b) and (c), two observations can be made. First, the
separation distance between each spectra is proportional to their intensity, and this ratio does not
change with different wavelengths.
Therefore, in this case, the intensity distribution is
wavelength independent. Second, the spectra from -12.5 to 0 degrees are close to each other.
As the angle is increased outside this region, the intensity decreases quickly. Hence, there exist a
range of angles where the light intensity is more uniform thereby exhibiting Gaussian
distribution.
However, this range is not centered at the origin (0-degree).
To check this
assumption, the intensity at the specific wavelength of 254nm versus the angle is plotted and
presented in Figure 3.12.
2
x 104
Bulb 6312-50 (1 mm aperture) Intensity versus Angle
I
+
100 mm away from bulb
+
1.8
+
±
+
95%
. .~
1.6
'
E
'AI
:.....
90%
.....-
...
+
1.4 kband Iw rdth
15 d grees
......
C4
.
.....
.
+
- .....
..
I
0.8
V.0 -
-20
- -..
..
..
-.
..
-15
-A - ..
........-..
..
--...
...
.
-..
-..
-10
......
-.
.....
.....
...
-5
0
5
Angle from certer(degrees)
10
15
20
Figure 3.12: Lamp intensity at 254nm versus the angle.
As seen in Figure 3.12, the maximum intensity is at -5 degrees, therefore in the system, the
lamp should be rotated counterclockwise 5 degrees. Also, a section of 15 degrees centered at -5
degrees will ensure 95% uniformity of the lamp output.
36
Figure 3.13 shows the comparison between the two different lamps. Using the same set-up
as Figure 3.11a, the maximum intensity between 200nm and 220nm versus the angle was
recorded for each lamp at three different distances (50mm, 70mm, and 100mm).
Lamp Comparison
L6311 (0.5mm aperture) L6312 (1mm aperture)
x 10
Distance flrom lamp
E3
X
5
L6312, 100mm
0 L6312, 70mm
A L6312, 50mm
E
a
0
+ L6311, 100mm
-6 L6311, 70mm
A
e~j 4
0
x
_
__
__
0
0
C
L6311,
50mm
+
0
A
Z%
A
0
X0
A
00
+
1
-50
-A
_
-40
-30
-20
-10
0
10
_-A-
20
30
40
Angle from center(degrees)
Figure 3.13: Lamp intensity versus the angle for each bulb.
This graph shows that both lamps have an off center peak around -5 degrees. As expected,
the 0.5mm aperture lamp is brighter and has a sharper peak than its 1mm counterpart. This
means that the 0.5mm aperture lamp should be used if light throughput is important, whereas the
1mm aperture should be used to provide a more uniform beam when using the 15-degree
bandwidth. For the first prototype, the 1mm aperture is used since beam uniformity is more
important for molecule visualization.
37
3.4.4
Different Approaches to Filtering
The deuterium lamp that was selected emits a broadband spectrum.
filtered so that the user can select a specific wavelength.
Its output needs to be
Two different filtering methods,
rotating filter wheel and diffraction grating, were identified and will now be presented.
Filter wheels are commonly used to select a specific wavelength beam. Band-pass filters that
only transmit light of a specific wavelength are attached to a wheel as seen in Figure 3.14. The
wheel is rotated about "axis 1" to bring the desired filter in the light path.
Lens
Lens
0
Mirror
Axis
A
1
Deuterium
Lamp
185nm-400nm
2xis
Filter Wheel
Figure 3.14: Filter wheel approach.
Band-pass filters are simpler in design and have a higher throughput than most other filtering
methods. However, this design's disadvantage is that only a finite number of wavelengths can
be selected. One way to expand the application of this design is by taking advantage of the fact
that the transmitted wavelength through a filter varies with the incidence angle of the light.
Therefore by rotating the filter wheel around "axis 2", the transmitted wavelength shifts towards
shorter wavelengths with increasing angle. The relation between the angle and the transmitted
wavelength is [10]:
A = AO 1
-n-
sin20
(3.1)
Where
A
is the wavelength at angle of incidence,
Xo is the wavelength at normal incidence,
O is the angle of incidence,
no is the refractive index of external medium,
neff is the effective refractive index of filter.
38
Variation of wavelength with incident angle
160
140
-+-
E 120 -100
C
- - -
---
-
-
- -
--
260rn
134.39
-- 2350nm
80
"I1go
60 40
3524
20
3.97
0
5
1.-0
10
14.87
15
20
25
30
35
Degrees
Figure 3.15: Variation of wavelength with incidence angle.
Figure 3.15 shows the variation of wavelength with the incidence angle for the UV region
(260nm) and for the IR region (2350nm). The transmitted light intensity decreases sharply after
approximately 25 degrees; therefore the maximum variation in the UV region is only 10nm.
This means that about 20 filters would be required to cover the 200-400nm range desired. In the
infrared region however, a variation of 95nm is possible at 25 degrees. This difference between
the two regions of the light spectrum is because the variation at a given angle is proportional to
the wavelength. In summary, this method could only be applied to an application in the infrared
but not in the ultraviolet range. Another method is therefore necessary.
The second filtering method consists of using a diffraction grating to spatially separate the light
into its monochromatic components. A schematic of a possible design is shown in Figure 3.16.
Rotates to
change focused
wavelength
Entrance
Slit
Lenses
Concave
Holographic
Grating
Exit Slit
Deuterium
Lamp
190=-400=
Shutter
Collimated
Output
Figure 3.16: Diffraction grating approach.
39
Collimating
Reflector
A brief overview of the diffraction grating concepts is presented below. For a detailed
explanation, one should consult appropriate literature such as reference [11].
A typical diffraction grating consists of a substrate with a large number of grooves
(>600lines/mm) ruled on its surface as seen in Figure 3.17.
grating normal
incident light
0
diffracted light
1lg
diffracted
light
d
Figure 3.17: Diffraction grating [11]
The incident light on the grating surface is diffracted into discrete directions. The grating
equation, which governs this diffraction, can be written as:
mX = d (sin a +sinfi)
(3.2)
where
m
A
d
a
,8
is the order of diffraction (an integer),
is the wavelength,
is the groove spacing,
is the incidence angle,
is the diffraction angle.
A common application of diffraction grating consists of changing the wavelength by rotating
the grating about its axis, with the incidence and diffraction light direction remaining constant.
The angles a and /8 change as the grating is rotated, but the difference between them remains
constant. This angle is called the deviation angle, 2K:
(3.3)
2K = a -8 = constant.
The grating equation can be rewritten as
mA = 2d cos K sin 0,
(3.4)
S= (a + 8)/2 = scan angle,
(3.5)
where
40
Equation (3.4) is required to design the mount for the grating. It shows that the diffracted
wavelength at the output slit is directly proportional to the sine of the angle
grating rotation angle.
#,
which is the
This relation is the basis for the grating driving mechanism seen in
Figure 3.18.
axis of grating rotation
(out of page)
grating
screw
Figure 3.18: Grating driving mechanism. [111
The relation between the arm length L, the distance X, and the angle
# is:
sino =-.
L
(3.6)
Finally, the grating equation can be written as
X
mA = 2d -cosK.
L
(3.7)
Equation (3.7) shows that as the distance X increases, the diffracted wavelength A increases
linearly with X.
It is important to note that more than one combination of m and A will satisfy the equality of
Equation (3.7). For example both m=1, A=A and m=2, 2=2/2 are valid solutions. This means
that at a given angle, there is overlapping of diffracted spectra as seen in Figure 3.19.
grating
normal
0
m = +2
300
nm
in = +1
200
nm
600
nm
incident
light
100
nm
400
nm
200
nn
Figure 3.19: Overlapping of spectral orders [111
41
Since usually only the first spectral order is desired, the higher orders may need to be
blocked by using some order-sorting filters.
Even though the diffraction grating system has a smaller light throughput than the filter
wheel, its ability to scan the entire spectrum makes it the first choice for the variable light source.
Since the deuterium lamp only produces a useful spectrum from 200-400nm, no order
overlapping will occur.
3.4.5
Different Diffraction Gratings
There are different ways of mounting diffraction gratings. They can be grouped into two
categories: plane grating mounts, and concave grating mounts.
Plane gratings need a collimated incident light to disperse the light by wavelength. Optics
cgrating
such as mirrors or lenses are needed to collect and focus the beam. Figure 3.20 shows the four
different types:
collimator
entrance slit
entrance slit
exit slit
exit slit
grating
camera
a) Czerny-Turner mount
mirror
b) The Ebert- Fastie mount
grating
entrance slit
mirror
exit sit
exit slit
grating
mirror
c) The Monk-Gillieson mount
d) The Littrow monochromator mount
Figure 3.20: Plane grating mounts. [11]
42
The Czerny-Turner (Figure 3.20a) is the most common type of plane grating mount. The
incident light is collimated with a mirror, and the diffracted light is focused with another mirror.
Since the light reflecting off the grating is collimated, there is no aberration introduced by the
grating, and the spectrum is always in focus at the exit slit. The mirrors however, introduce
astigmatism and spherical aberration. The Ebert-Fastie mount (Figure 3.20b) is similar to the
Czerny-Turner, with the difference that it uses one large mirror for both collimating and focusing
the beam. This mount is not often used because aberration and stray light are difficult to control.
In the Monk-Gillieson mount (Figure 3.20c), a mirror is used to converge the beam onto the
grating.
Since the incident light on the grating is not collimated, wavelength dependent
aberrations are present and the beam is not always in focus at the exit slit. To counteract this
problem, the grating needs to be rotated about an off-center axis and the angles of incidence and
diffraction should be kept to a minimum. This type of grating is popular for low-resolution
applications since it is the cheapest type of mount available. Finally, the Littrow monochromator
mount (Figure 3.20d) is used primarily in laser tuning applications. The incident and diffracted
light beams have almost the same direction. The entrance and exit slits are slightly offset, which
introduces out-of plane aberrations.
On the other hand, concave gratings combine the function of optical imaging and wavelength
diffraction in one component, such that external optics are not necessary. See Figure 3.21.
incident light
source point
A
B
image point
diffracted light
Figure 3.21: Concave grating mount [11]
43
The most common type of concave grating mount is the Seya-Namioka, which scans the light
spectrum simply by rotating the grating about its axis. This grating offers two main advantages
over plane gratings.
It has a lower aberration at the designed wavelength, and a higher
throughput since no other optic component is needed. It is the preferred type of grating mount
for UV spectroscopy and therefore will be used in the design of the light source.
The Richardson grating laboratory offers a wide variety of high quality concave gratings. To
maximize throughput, the grating needs to be as large as possible and have the largest number of
grooves per millimeter. The grating coating and blazing wavelength affects its efficiency and
consequently also influences the system throughput. For the proposed light source, two possible
concave holographic gratings have been selected. Their characteristics are listed in Table 3.2:
Grating
Grooves
per mm
Imaging Range
(nm)
Blaze
wavelength
Dispersion
(nm/mm)
Free Aperture
(mm)
Input
F/#
(nm)
1
1200
200-800
250
8.5
30 * 30
3.6
2
1350
190-700
250
8.0
p40
2.3
Table 3.2: Selected concave holographic grating from Richardson grating laboratory.
The Richardson grating laboratory only manufactures its concave grating on demand, and
thus 8 weeks lead time is required for most of their gratings. To avoid the wait, an off-the-shelf
monochromator model 9030 from Sciencetech, Inc was purchased. This offers two advantages
that simplify the light source design. First, the grating is similar to #1 in Table 3.2 except that it
has a dispersion of 8nm/mm and an F/3.2. The F/# is the ratio of the lens focal length to the lens
aperture diameter. Second, the grating is already mounted, and the sine drive to rotate it is
included. Some major modifications of the monochromator were made to adapt it to the design
and will be explained in Section 3.4.7.
44
3.4.6
Optical Components
Now that the deuterium lamp and the concave diffraction grating filtering method have been
selected, the optics to focus the lamp beam into the entrance slit and to collimate the output from
the exit slit are investigated. See Figure 3.22.
Rotates to
change focused
wavelength
Entrance
Slit
Lenses
Concave
Holographic
Deuterium
-Emp
GratingA-ExtSi19=40
Shutter
Collimated
Output
Collimating
Reflector
Figure 3.22: Optics set-up to focus and collimate the light
The collimator is described first since it is the simpler of the two systems. The diffracted
light coming out of the exit slit of the grating mount has a F/3.2. It needs to be collimated and
redirected at a 90' angle for the output beam to be vertical. An off-axis paraboloidal reflector
with a diameter of 2.5 inches and a focal length of 119mm from Oriel Instruments was first
selected, since it has a F/3.3 and can both collimate and redirect the light vertically. However,
due to the high price and the required lead-time of 6 weeks, another approach was adopted. A
concave reflector, model 7292, with a focal length of 125mm and a F/3.2 from Oriel instruments;
and a square mirror model K45-340, 75mm, UV coated from Edmund Industrial Optics, are used
instead. This system is composed of two elements but it has the advantage that each component
is mounted on a separate minimount that allows for fine tilt adjustment, which is crucial for
calibration of the system.
The incident light on the grating has to be focused to a F/3.2. Three different methods, using
lenses, reflector, and direct illumination have been designed. After presenting each method, the
test results for each are shown and explained.
45
The first method uses two fused silica plano-convex lenses from Coherent Inc. to match the
F/# of the incident light with the F/# of the concave grating. See Figure 3.22. The first lens,
with a focal length of 50.8mm, collimates the light coming from the deuterium lamp, whereas the
second lens with a focal length of 76.2mm, focuses the light on the entrance slit with the correct
F/#. This method has the advantage of maximizing the incoming light, however the lenses
introduce chromatic and spherical aberration. Chromatic aberration is the chromatic spreading
of light. Each wavelength is focused at a different point along the optical axis, because the index
of refraction of all optical materials varies as a function of wavelength. Spherical aberration is
the distortion of the image and the creation of a blur at the focal point because different radial
sections of the lens focus at different points. To minimize spherical aberration in this set-up,
lenses that are twice as large as the light beam diameter are used. Moving the lens slightly
toward the focal point also reduces the aberration by averaging the error across the entire lens
instead of just the outer rings.
Deuterium
Lamp
Rotates to
change focused
wavelength
Focussing
Reflector
Concave
Holographic
Grating
Entrance Slit
Exit Slit
Shutter
Collimated
Output
Collimating
Reflector
Figure 3.23: Light source set-up using a reflector.
The second focusing method consists of using a square reflector model 7295 from Oriel
Instruments with an 86mm focal length as shown in Figure 3.23. This system differs from the
lens set-up, as a concave reflector replaces the lenses. This eliminates chromatic aberrations.
However, off-axis aberrations are now introduced.
46
Rotates to
change focused
wavelength
Deuterium
Lamp
Concave
Holographic
Grating
Shutter
Collimated
output
Collimating
Reflector
Figure 3.24: Light source set-up with direct grating's illumination.
The third and last method is shown in Figure 3.24.
It consists of eliminating all of the
focusing components and placing the deuterium lamp at the entrance slit position. This approach
might seem to be against all the good practices of an optics system design. However, due to the
special characteristics of the designed system, this method is feasible and its advantages could
exceed its disadvantages. As seen in the Section 3.4.3, the deuterium lamp has a small aperture
(0.5mm for the L6311 and 1mm for the L6312). Therefore the lamp aperture can replace the slit.
Moreover, the acceptance cone of the grating, with its base at the grating surface and its peak at
the entrance slit, has an angle of 180. As seen in Figure 3.12, the lamp has 95% illumination
uniformity for an angle of 15*. Consequently, by placing the deuterium lamp at the entrance slit,
the light incident on the grating is at least 92% uniform. This has been estimated from the data
of Figure 3.12. By avoiding the focusing optics, chromatic and spherical aberrations should be
minimized. However the light dispersion is expected to be larger.
Now that each of the three methods, using lenses, reflector, and direct illumination, has been
presented, the experiments to compare them and their results will be presented.
parameters are of importance in the comparison of the different systems:
"
the uniformity of the illumination at different wavelengths,
"
the bandwidth of the spectrum,
"
the shift of the spectrum in the beam profile.
47
Three
To test the uniformity of the illumination, the light output was visualized at eight
wavelengths (200,230,260,290,320,350,380,and 400nm) by taking a snap-shot picture using the
CCD camera. See Figure 3.2a for the imaging set-up diagram. Since in the final application the
camera will be taking a picture of a gel placed above the light source, the camera was focused on
a plane just above the light to mimic its future application. A 3D-mesh plot of the results is
shown in Figure 3.25, 3.26, and 3.27.
To find the bandwidth and the spectrum shift throughout the beam, a spectrum of the output
beam was recorded at different locations in the beam and at different wavelengths by using the
spectrometer. See Figure 3.2b for the spectrum analysis set-up diagram. The fiber optic was
placed vertically above the light source to also mimic its future application.
The results are
shown in Figure 3.28.
The L6312 lamp (1mm aperture) was used in this experiment for all three set-ups. The
entrance slit for the lenses and reflector set-up had a 1mm width and an 8mm height. The slit
width was selected to match the aperture of the L6312 lamp so that all three set-ups have the
same slit entrance size, and therefore can be compared under the same conditions. The exit slit
was first chosen to match the entrance slit for optimal throughput. However, after initial tests, it
was observed that the output intensity was so large that the detector always saturated. To solve
this problem, an exit slit of 0.5mm width was placed to reduce the output signal intensity. The
beam intensity was reduced by half, however the bandwidth of the output was slightly larger than
the value of 4nm/mm predicted by the grating specification. The grating dispersion and a
convolution of the input and exit slit width determine the bandwidth. In the set-ups, the entrance
slit was 1mm wide, the exit was 0.5mm wide, and the grating dispersion was 8nm/mm.
Therefore, the bandwidth for the lenses and the reflector systems should be around 6-7nm.
48
Set up with lenses at 200nm
Set up with lenses at 230nm
3000, .....-.--.2000,
-1000 , ...
3000
20001.
1000
-...
0
--
0
50
50
0
20
60
40
60
40
0 20
Distance 10 units = 9.3mm
Set up with lenses at 290nm
Distance 10 units = 9.3mm
Set up with lenses at 260nm
3000,
2000 ,b
1000
...
---..-..
3000,
50
40
0 20
Distance 10 units = 9.3mm
Set up with lenses at 320nm
U)
0
50
-...
- -..
....
0
--
20
60
40
Distance 10 units = 9.3mm
Set up Wth lenses at 350nm
3000
2000,-.---
-..
.. .. . ...
-
0
50
40
0 20
Distance 10 units
Set up with lenses at 380nm
3000
: 20001
4) 1000
-......
1000
040
50
3000
2000
-.....-.
0
= 9.3mm
20
Istance 10 units =9.3mm
Set up with lenses at 400nm
--
3000
-......
2000
-..
-....1000 ......-.
-....
-
0-
50
50
0
20
40
60
40
Distance 10 units = 9.3mm
Figure 3.25: Light source
0 20
60
Distance 10 units
illumination at different wavelengths for set-up with lenses.
49
=9.3mm
Set up with focusing mirror at 200nm
Set up with focusing mirror at 230nm
4000,
-
2000'....-
-
0
2000
0
u.. hfcsn
0
50
50
40
60
o a
40
0 20
Distance 10 units = 9.3r nm
Set up with focusing mirror at 260nm
60
Distance 10 units = 9.3mm
0 20
Set up with focusing mirror at 290nm
4000
4000
-...
2000,..---
2000 ,.
--
0
50
-
50
0
20
40
40
Distance 10 units = 9.3r nm
Set up with focusing mirror at 320nm
0 20
Distance 10 units = 9.3mm
Set up with focusing mirror at 350nm
---
4000,--
2000 ......-.-.
U2000,..
- --
0
50
--
4000,..
50
40
0
60
20
Distance 10 units
Set up with focusing mirror at 380nm
4000'...-Vi2000 .-
60
0 20
9.3mnm
40
Distance 10 units = 9.3mm
Set up with focusing mirror at 400nm
-
4000
---
---
2000,.--0 ... .. . ..
50
50
0 20
40
Distance 10 units
9.3p mr
0 20
Distance 10 units =9.3mm
Figure 3.26: Light source illumination at different wavelengths for set-up with reflector.
50
Set up with no focusing optics at 200nm
Set up with no focusing optics at 230nm
4000
20...,....-.
'2000
50
--
50
0 20
40
60
40
Distance 10 units = 9.3r nm
Set up with no focusing optics at 260nm
4000 ,.......-.
0 20
Distance 10 units =9.3mm
Set up with no focusing optics at 290nm
---...
4000
Wi2000,... -.--
60
I2000,.
0
50
50
0 20
40
60
40
Distance 10 units = 9.3r nm
Set up with no focusing optics at 320nm
4000 ......
--
wi2000,......--.
S 0'...- -50
2)
0 20
Distance 10 units =9.3mm
Set up with no focusing optics at 350nm
4000
-....
-.
2000
-......
0
--....
..
50
'6
40
60
60
40
0 20
Distance 10 units = 9.3mm
Set up with no focusing optics at 400nm
0 20
Distance 10 units = 9.3r nm
Set up with no focusing optics at 380nm
4000',.. . - -
4000
W2000,....---
i2000,-.-
50
50
0 20
40
Distance 10 units =9.3r
IM020
0 240
Distance 10 units
=9.3mm
Figure 3.27: Light source illumination at different wavelengths for set-up with direct illumination.
51
Maximum variation of spectrum location in the beam profile
Imm aperture, 0.5mm slit
6
4
x 10
Set-up with lenses
-- Beamt
x2
4-
profile
X1
.
3
----1- --.-----
-.
--. - -.--
--..
--
-.
2-
......... ...
.........
...-.--.-- . ----
.
.......
..
E
....
1
a)
U
245
40
250
255
260
265
270
275
280
28 5
Wavelength (nm)
6
x 10
Set-up with mirror
5
4
U)
C
4)
X1s
-..
- ....
....
..- -..
3 . .....
-........
...
.. ..
.
..............
.........-..
2
b)
1
0
240
6
245
x 10
250
255
260
265
Wavelength (nm)
270
4
275
280
285
Set-up with no optics
5
I
4
3
xo
- --- - --
2
-- - -- -
- ----- ---
-----.-.-- --- -- -
- -----.. ---
-- -- -- -- - -- -
2
- ..-..-
- -
---
-
-- - -
1
c)
0
24 0
245
250
255
260
265
270
275
-
280
Wavelength (nm)
Figure 3.28: Light source spectrum analysis for three different focusing set-ups
a) set-up with lenses, b) set-up with reflector, c) set-up with direct illumination.
52
285
Figures 3.25, 3.26, and 3.27 show the illumination from the three set-ups. The picture of a
ruler was taken to correlate the pixel number with the actual image size (mm). From the mesh
plot, some conclusions were drawn about the uniformity of each set-up.
For all systems, the intensity is wavelength dependent.
This is to be expected since the
deuterium lamp intensity is wavelength dependent as seen in Figure 3.11. Besides the lamp, the
optics and the detector efficiency also influence the final output intensity.
The set-ups can be arranged in order of increasing throughput: the lens system, the reflector
system, and the direct illumination system. There are two reasons for this behavior. First, there
are losses associated with each optical component that transmit or redirect the light. The more
elements a system has, the higher its losses. The lens system has three elements, two lenses and
a slit, to focus the beam; the reflecting system has two, the concave mirror and the slit; and the
direct illumination system has none. This correlates with the result observed in the figures. The
difference between the lens and the reflector system is further accentuated by the fact that lenses
inherently have a lower efficiency than reflectors. Second, because of their larger size, reflectors
can collect more light than lenses. The reason that the direct illumination system has a higher
throughput lies in the fact that the deuterium output is highly directional. As seen previously,
most of the beam intensity resides within a 20-25 degree cone. Since the acceptance cone of the
grating is around 18 degrees, almost all of the incident light reaches the grating.
As seen in Figure 3.25, the illumination from the set-up with lenses is not uniform. This is
mostly due to the spherical aberration introduced by the lenses, but is also due to misalignments
between the lamp, lenses, slit, and grating. Even though spherical aberrations were minimized
by using lenses twice as large as the beam diameter, they were still significant because of the low
F/# of the lenses used.
Using a reflector instead of lenses in the focusing system greatly improves the illumination
properties as shown in Figure 3.26. The light is more uniform and the profile of the illumination
is larger than the lens system. The flat areas at the top of the profile at 260, 290, and 320nm are
due to the detector saturation.
Figure 3.27 shows the results of the direct illumination system. It is observed that the beam
profile is larger than the lens and reflector systems. Most of the plots show that the detector
saturated due to intense light. However, some conclusions may be made. The part of the plot
53
that is not saturated suggests a uniform beam similar to the reflector system. The figures where
the detector has not saturated confirm this hypothesis.
It should be noted that these 3D mesh plots (Figure 3.25, 3.26, and 3.27) only give an
estimation of the illumination profile. The camera UV-lens system is focusing on a finite plane,
trying to image the beam at that plane.
However, since the beam is collimated, there is a
superposition of imaging planes, which induces errors in the illumination. Nevertheless, this
approach is sufficient for the purpose of comparing the different set-ups. For the final design, a
scanning approach using fiber optics is implemented to give an accurate measurement of the
output beam profile.
In conclusion, the illumination test illustrates that the profile of the lens system is not
uniform, whereas the reflector and direct illumination systems have relatively uniform profiles.
Also, the direct illumination set-up has a much higher throughput than the other two systems.
Another important characteristic of the light beam is needed to compare the different set-ups.
Figure 3.28 shows the results of the spectrum shape and shift at three different locations in the
beam. Position XO is the center of the beam whereas position X1 and X2 are respectively the
right and left side of the beam profile.
The spectrum intensity values at X1 and X2 were
multiplied by a factor so that their peak intensity matched the XO spectrum peak intensity. The
parameters of interest extracted from the data in Figure 3.28 are summarized in Table 3.3.
Set-up system
Bandwidth (nm)
Shift in spectra (nm)
With Lenses
With reflector
Direct Illumination
7.0
7.0
15
6.0
4.6
0.7
Table 3.3: Results of the spectrum bandwidth and location test.
The results of this test confirm our predictions.
The system with lenses has a small
bandwidth of 7nm, but also the largest shift in the spectra location (6nm). This is mainly due to
chromatic and spherical aberrations introduced by the lenses. The shift in spectra is of the same
magnitude as the bandwidth.
Therefore as seen in the figure, the spectrum at X1 and the
spectrum at X2 are far enough from each other than there is almost no overlapping between
them.
The reflector system exhibits the same characteristics as the lenses system. It has a small
bandwidth of 7nm and also a shift of 4.6nm. The spectra locations do not vary as much since
only spherical aberrations are present.
54
The direct illumination system on the other hand has only a 0.7nm shift in its spectra
locations along the X direction. In the direction perpendicular to X, a shift of up to 2nm was
observed. This large improvement comes at the cost of having a larger bandwidth of 15nm.
The shift in the spectra observed in the lens and reflector systems is not acceptable for this
application because the light output will be of a different wavelength than expected and this
could lead to large errors in measurements. Therefore, the direct illumination method will be
used in the variable light source. The bandwidth of 15nm is within the specification for imaging
the gel. However, the shape of the spectra for the direct illumination set-up in Figure 3.28 is not
perfect and suggests that this method could be optimized. Possible optimizations are explained
in Section 3.4.8.
3.4.7
Final Design and Lamp Characteristics
Now that the main components of the light source have been selected, namely the deuterium
lamp, concave holographic diffraction grating, and collimating optics; the final design of the
light source and its characteristics are presented.
Slit
Electronic
shutter
Concave grating +
motorized assembly
Collimating
optics
Output
window
Deuteurium
lamp
Heat sink
Figure 3.29: Proposed light source
55
The detail and assembly drawings for the light source are found in Appendix C1 and C2. As
seen in Figure 3.29, the light source is separated into two compartments, the lamp and the
collimating section. The lamp section contains the deuterium lamp, the diffraction grating and
its motorized assembly, the cooling fan, and the electronic shutter.
The collimating section
includes the exit slit, the collimating reflector, and the 45-degree mirror.
The main optical component mounts, lamp, grating, and focusing optics, permit a full range
of motion to enable the user to precisely adjust the component position and rotation.
The deuterium lamp has four degrees of freedom. It can be rotated through its vertical axis,
and can be displaced horizontally and vertically. Also, the lamp holder allows for a change of
the lamp without losing the positional settings. The deuterium lamp has a 30W output, and
therefore a cooling system is needed to ensure that no overheating occurs. Overheating would
result in poor performance and shorter life of the lamp. On the cover inside the enclosure, a
small inverted heat sink is mounted above the lamp to prevent local heating of the cover. A
small fan with a filter is mounted inside the frame to recirculate the air inside the lamp side of
the light source. The fan and exit holes are placed in such a way that the filtered air passes by
the lamp, the heat sink, the stepper motor, and then exits behind the motor.
The diffraction grating has five degrees of freedom. Its mount is based on the Sciencetech
9030 monochromator.
Of the monochromator, only half of the base was kept because it
contained the sine drive and an adjustment knob with a wavelength display. In the original
monochromator, a mirror redirected the diffracted beam toward the exit slit.
A clockwise
rotation of the knob resulted in a clockwise rotation of the grating and an increase in the output
beam wavelength. Since in our set-up there is no mirror between the grating and the exit slit, a
clockwise rotation of knob would result in a decrease in the output beam wavelength, which is
the reverse of the counter display. To correct this problem, the grating needs to rotate in the
opposite direction when the knob is turned clockwise. This was accomplished by inverting the
base, and mounting the grating and its holder on the opposite side of the shaft. In this set-up, a
clockwise rotation of the knob results in a counterclockwise rotation of the grating, and as
desired, an increase in wavelength.
A stepper motor and flexible coupling are mounted on the side of the diffraction grating base
to automate the grating rotation via a computer. Turning the knob rotates the grating so that a
different wavelength is focused on the exit slit. Each turn of the knob corresponds to a 100nm56
wavelength scan.
The motor was selected to have 200 steps per revolution, resulting in a
resolution of 0.5nm in the focused wavelength. A miniature switch is mounted under the base to
allow for an automatic wavelength calibration when under computer control. A power switch on
the back panel allows for switching between manual and motorized operation of the grating
rotation.
When the light source is used for imaging a gel, an electronic shutter is needed to control the
exposure time. The spectrometer contains a shutter. An identical one was purchased, a Prontor
Magnetic 016 shutter from Schneider Optics, as to use the same controller for the light source
shutter. However, using the same controller means that the light source shutter needs to be
removed when using the spectrometer, since the controller can only operate one shutter at a time.
There is a controller inside the CCD camera that could be used to control the light source shutter,
but this would require the machining of some the CCD components to allow for the connection
to be made.
The exit slit came with the monochromator and is mounted on the inside wall. It can be
easily replaced with a different size slit. If the slit width needs to be adjusted automatically,
motorized slits are widely available and can be adapted to the light source. The collimating
reflector and the 45 degree mirror have five degrees of freedom for easy calibration.
To minimize stray light in the system, the inside wall needs to be coated with a matte black
paint. However, optics researchers at Acton Research warned that even UV resistant paint would
deteriorate quickly under the intense direct UV illumination of the deuterium lamp. The paint
would chip away from the wall, and paint dust particles would start depositing all over the optics.
This would seriously reduce the lamp performance and would therefore be unacceptable.
Therefore, the walls in the lamp section of variable light source are left bare. The light reaching
the collimating section is of much lesser intensity, and thus the walls in that section are painted
with an ultra matte black paint.
The effects of leaving the walls bare in the lamp section were measured to check their
importance. The experiment was performed in two parts. First, the reflectivity of an aluminum
plate in the UV region was compared to a mirror and to a black wall. Second, the influence of
the reflection in the light source was measured by blocking the acceptance cone of the diffraction
grating, and measuring the output spectrum as seen in Figure 3.30.
57
Concave
grating
Deuterium
Baffle Bafflelamp
5'A
Exit Slit
- ----------------Reflected light
Unwanted
output
Collimating
reflector
Figure 3.30: Set-up to measure unwanted reflected light.
Wall type
Reflectivity relative
to straight light (%)
Mirror
76.9
Bare aluminum
Painted Aluminum (black)
9.7
0.3
Table 3.4: Different surface reflectivity.
The reflectivity at 260nm of different surfaces relative to direct light is shown in Table 3.4.
The reflectivity of bare aluminum is 9.7% and is significant. However as seen in Figure 3.31,
the test measuring the influence of the bare aluminum walls in the light source reveals that they
have no effect on the output signal.
x 104 Spectrum without the blocking baffle
.-..-- -
-.
..
.
..
5 -- ---.
-
Spectrum with the blocking baffle
140
- -.....
--
130
120
3--
-
1 10
- -. --..- -- -
-- -
100
90
1
0
200
300
400
80
500
Wavelength (nm)
------...-.-.-.-.----
200
300
.---..-.---- .---- .---- .-..--
400
Wavelength (nm)
Figure 3.31: Influence of bare aluminum walls on the output signal.
58
500
Now that the final design has been presented, the light beam output power and profile will be
precisely characterized.
By using the spectrometer with the fiber optic, the output power
throughout the spectral range was recorded and is shown in Figure 3.32. In the figure, some
characteristic spectra are shown at selected wavelengths of 210,260,310,360,and 400nm.
Variable light source Power output
X 10,
0r
4.5
- ......................
4
...
..........
..........
3.5
3
-
-..-....
......
-.
a) 2.5
-..
C
2
-- -
-.
-...
--
-
-..-.-.---
1.5
1
0.5
U
190
-2.
210
260
310
360
400
450
wavelength (nm)
Figure 3.32: Output power of the variable light source.
A program was used to control the XY stages and record the spectrum of the light at 260nm.
The spectrum was recorded every 2mm throughout a square area of 40mm length. The resulting
400 spectra were analyzed to extract the wavelength and the intensity of the maximum peak.
Mesh plots of the results were created and are shown in Figures 3.33 and 3.34. The light source
output beam has an area of around 32mmx32mm. Figure 3.34 shows that there is a small shift in
the peak location. On the side the picture, the surface is rough. At those locations, the data was
just background noise because there was no illumination. Consequently, the Matlab program
used to extract the maximum peak wavelength generated random values at these locations.
59
Mesh plot of Light illuminaton at 260nm obtained using scanning
x10
6
5
:3-
0-
20
105
15
10
15
10
X,Y Distance 1 unit
05
20
2mm
Figure 3.33: Illumination profile of light source at 260nm.
Wavelength at the peak throughout the illuminated profile
0.
0
290
--
-I
----
.
280,270,s
260s.-
240
.
5
10-
-920
15
15
10
20
5
25
0
Distance 1 unit =2mm
Figure 3.34: Peak location throughout the beam.
60
3.4.8
Recommendations and Future Work
In this section, three possible improvements of the light source are described.
The first
improvement consists of increasing both the throughput of the system and the output beam size.
The deuterium lamp has large directivity and using a larger grating would be more effective.
This should be combined with characterizing the effect of using different slit widths and heights.
In all the experiments, the slits had a height of 8mm. A longer slit would increase both the light
throughput and the beam size, if the output beam in the exit slit plane was long.
The second improvement consists of reducing the bandwidth of the output beam.
As
mentioned previously, the shape of the spectrum in the final design shown in Figure 3.28c is not
a true Gaussian, and therefore, could be improved. The shape of the spectrum is possibly the
addition of three different spectra as shown in Figure 3.35.
Sum of the spectra
k/
k0
U
Figure 3.35: Possible Gaussian addition to explain the spectrum shape
The light path helps us to understand the source for the extra peaks on the side of the
spectrum. Figure 3.36 shows a schematic of the light bulb and its light path.
Unwanted light
Diffraction
Deuterium lamp
grating
Desired light
Figure 3.36: Deuterium lamp light path
61
As was explained previously, the incident light on the grating needs to come from one point
source for optimum performance of the system. The deuterium lamp has a small aperture, and
ideally, its light output path should be a cone. However, the lamp is enclosed in a cylindrical
glass bulb, and as seen in the drawing of Figure 3.36, this induces some of the light to be
refracted toward the grating. Since this light is coming from a different point than the desired
light, it will diffract on the grating at a different angle and appear in the output beam with a
different wavelength.
A simple experiment was performed to check this assumption. The experiment consists of
placing a baffle between the deuterium lamp and the grating on either side as shown in Figure
3.37.
This would eliminate some of the undesired light and reduce the bandwidth of the
spectrum.
Left side
baffle
Deuterium
Lamp
Concave
Holographic
Grating Grat---
Exit Slit
Shutter o
Collimated
Output
Collimating
Reflector
Figure 3.37: Direct illumination with a baffle to restrict the light reaching the grating.
The results of this experiment are shown in Figure 3.38. Placing the baffle on the left side of
the lamp results in the thinning of the right side of the spectra with no effect on the location of
the center peak. See Figure 3.38a. This is similar to removing the peak corresponding to X2 in
Figure 3.35. Repeating this experiment with the right side yields the same result except that the
left side gets thinner. See Figure 3.38b. It may be concluded that the spectrum for the direct
illumination set-up is possibly an addition of three spectra as seen in Figure 3.35 and is caused
by the deuterium lamp.
62
Baffle test on direct Illumination Set-up
H
Left baffle
A x10,
I
4
Shift in spectrum with Direct
illumination and baffles
X 10
No baffle
3
3
...
.....
.. ......
..
...
.....
.....
P eam Profile
Left baffle
.
......... .......
...... .......
X0
2
.........
1
x
.
X2I
*X
... . . .
X1
2
I
d)
a)
240
250
260
270
280
M
240
290
250
Wavelength (nm)
4
x 10
4
270
260
Wavelength (nm)
280
Spectrum at different wavelength
Right baffle
x 10e
for set up with baffles
-
..............
No baffle
Right bafife
..........
3 ..-.
.....
...
--.
....
3
...............
~ ~ ..
...
~ ..
~ ..
~....
~ ..........
~ --
2
-.-.---..
.-..--.---..
-
- -- --
- ---....
-....
-- ...-.....
1
e)
b)
240
4
250
260
270
280
Wavelength (nm)
A
290
200 220 240 260 280 300 320 340
Wavelength (nm)
Both baffles
x 10
--
No baffle
Both baffles
3
..
...
....
.. . .
Fiber
optic
... ..................
U)
(2 -
..
..
......
. .....*
. .. ... .........
.. .
1
1
....... ......
. .....
....
...
..
X2
X0
c)
0
24 0
250
260
270
280
290
Wavelength (nm)
Light Output Beam
Figure 3 38: Results from testing the influence of the baffles on the direct illumination set-up.
63
Xi
This experiment suggests that adding a baffle on each side of the light source reduces the
bandwidth of the direct illumination method. A direct illumination set-up using baffles was
developed and tested. As seen in Figure 3.38c, the bandwidth was reduced from 15nm to 9nm,
however the illumination profile was much smaller and the spectrum location shifted at different
locations in the beam. See Figure 3.38d. Due to the obstruction by the baffles, the grating was
not entirely illuminated. This resulted in the smaller profile obtained. The shift of the peak
location is much larger than the one measured without baffles, ± 6.5nm versus ± 0.35nm, and is
possibly paused by the interference patterns introduced by the baffles. Despite this spatial
wavelength shift, the spectrum shape was consistent for different wavelengths. See Figure 3.38e.
Now that it has been demonstrated that the lamp is most likely the origin of the problem, two
different approaches could be taken to solve it and should be investigated. The first one consists
of conducting more research on the effect of baffles and could result in an improved performance
of the current light source. The second approach consists of eliminating the problem by using a
different lamp. Hamamatsu offers another type of deuterium lamp called projecting type lamp as
shown in Figure 3.39.
(External view)
(Construction)
Lood www8IL
Figure 3.39: Projecting type deuterium lamp.
(Courtesy of Hamamatsu Photonics)
The projecting type lamp has a more uniform transmittance due to the plane glass and a
smaller directivity. Using this lamp could reduce both the spectrum bandwidth and the shift in
the peak location. However, the projecting type lamp requires a different power supply and a
different, more complex, lamp mount.
The third improvement consists of increasing the wavelength range of the light source into
the visible region of the spectrum by adding an halogen lamp to the system. The halogen lamp
would be placed behind a see-through Hamamatsu deuterium lamp. The light of the halogen
lamp would pass directly through the deuterium lamp. Some order sorting filters need to be used
in this design to eliminate spectra overlapping.
64
3.5 Cutting Tool
Once the bands are identified in the gel using the variable light source and the CCD camera, the
desired bands need to be excised and stored. An automated cutting tool designed specifically for
this application is presented in this section.
3.5.1
Specifications
The following are the requirements for the excising device:
" Excision of a band from an agarose or polyacralymide gel.
*
Expulsion of the band in a storage vial.
*
Extraction of band of varying rectangular shape,
5-10mm width, .5-5mm length, and 1-10mm thickness.
*
No cross contamination of samples between different cuts.
A device that can meet all these requirements is difficult to design. The fact that the gel and
bio-molecules it contains are fragile and temperature sensitive makes this task even more
challenging.
3.5.2
Existing Designs
The process of excising the desired band out of the gel is currently performed manually. The
operator locates the desired band by looking at the gel on a transilluminator. Then using a razor
blade, the band is cut and extracted. This process is neither repeatable nor accurate. Since the
operator is exposed to UV light and is handling hazardous chemicals with a sharp object, health
concerns are also a major issue.
An automated process can solve these problems and is clearly needed. At the time of the
design of the excising system, no automated system was available commercially. However, BioRad Laboratories has recently developed an automated spot cutter. The Bio-Rad system has
similar features to the one proposed in this thesis, namely it excises protein spots from a gel
using a mechanical cutter and deposits them on a microtiter plate. The specifications for the BioRad machine are listed in Appendix D4. There are three main differences between the two
systems:
65
1.
The Bio-Rad system was specifically designed for excising protein spots from a 2-D
electrophoresis gel. It employs a rotating circular blade to cut the gel. The cutting tip
has a round shape because in two-dimensional gels, the samples' shape is round. The
gels used in the purification of bio-molecules are usually one-dimensional, and the
bands have a rectangular shape. In contrast, the proposed system utilizes rectangular
cutting tips. Cutting tips of any shape can be mounted on the holder of the designed
machine, whereas the Bio-Rad machine has to utilize round tips because of the
rotating cutting motion.
2. The Bio-Rad system uses a Imm-diameter cutting tip that needs to be manually
mounted on the machine by the operator. If a large spot needs to be excised, then
multiple cuts are necessary.
Since a round cutter is used and as specified in the
instructions, overlapping the cuts is not recommended, the samples between the cut
circles will not be extracted. On the other hand, the proposed system automatically
loads different sized cutting tips depending on the size of the band to be excised.
Since the cutter shape is rectangular, multiple cuts next to each other will fully extract
the desired sample.
3. In the Bio-Rad system, the cutting tip height needs to be calibrated manually by the
operator to ensure that the gel is cut to the full depth.
This calibration is not
necessary in the proposed design because, as explained in Section 3.5.4, a feedback
system senses when the cutter touches a hard surface. This acts also as a safety
feature since the cutter will stop if an obstacle is encountered.
3.5.3
Possible Designs
In the design of the automated cutting system, the cutting method was first selected because the
rest of the design is dependent on it. The cutting device has to excise a band out of an agarose or
polyacrylamide gel. The gel has the consistency of gelatin. It is flexible yet brittle. Figure 3.40
shows different possible methods to cut the gel:
66
a)
e)
T
o
d)
c)
\b)
g)
h)
Figure 3.40: Potential design of the cutting device using a:
a) stamp, b) scoop, c) cutting blade, d) roller, e) wire, f) vibrating wire, g) air or water jet, h) laser
The first method consists of using a stamping device as shown in Figure 3.40a. This device
can excise the gel in one step, but has a fixed size, so different cutting tips of various sizes are
needed. The scooping device shown in Figure 3.40b can excise a band of fixed width and
varying length. A different cutter is needed to cut the end pieces of the band. All the other
designs can cut a band of any size, however a secondary operation is required to extract the band
from the gel. Figures 3.40(c) and (d) show respectively a blade cutter and a cutting roller. The
roller uses a serial process to cut the gel whereas the blade uses a parallel one. This means that
less force is required to cut the gel for the roller system than the blade. However, the roller will
make a longer cut than the desired band size because of its round external shape. Another
possible design consists of a thin wire passing between two very thin tubes as shown in Figure
3.40e. This design has several problems. It is difficult to obtain a high tension in the wire
because the tubes holding the wire cannot support a large bending moment. The tube size will
cause indentations at the corners of the cut that could result in an incorrect band shape. The
subsequent design shown in Figure 3.40f consists of a sharp wire vibrating in the ultrasonic
range. The high frequency vibration will create a crack that will propagate in front of the sharp
edge of the wire. It has the advantage that the wire stays clean because the high vibration will
prevent contaminants from sticking to the blade.
67
All the methods stated previously involve
direct contact between the cutter and the gel. Two other methods, laser and air/water jet cut the
gel without being directly in contact with it. See Figure 3.40g,h. These have the advantage of
eliminating cross contamination between different cuts. The laser cuts a material by sublimating
parts of it, and consequently the gel near the cutting point is heated during the process. The gel
is heat sensitive; it shrinks significantly when heated. Moreover, this heat could damage the
biological sample contained in the gel. These issues are especially important for thicker gels.
Therefore, a cooling method is necessary if a laser is used to cut the gel. The water and air jet
methods involve sending high-pressure jets onto the gel to cut it. Since the gel is placed on a flat
surface, the cutting fluid is trapped under the gel. This could cause the gel to move and some of
the biological sample to be washed away from the gel.
From the methods presented, the mechanical stamping arises as the method of choice. It has
the advantage of excising (cutting and extracting) the desired band in one step. It is also the
simplest method, which means that it is more reliable and less expensive.
3.5.4
Design of the Cutting device
Figure 3.41 shows the final design of the cutting device.
Assembly and detail drawings are
presented in Appendix Dl and D2 respectively.
The cutting device is composed of four subassemblies, main base, motor base, cutting tip
holder, and the cutting tip. The operation of the cutting device is as follows. During the excision
part, a band that needs to be excised is placed under the cutter. The motor base and the cutting
holder start moving downward. Once the band has been cut and the end of the cutting tip has
reached the gel tray surface, the cutting tip and its holder stop moving. However, the motor base
keeps moving down until the cutting tip holder activates the limit switch attached to the motor
base. Then, the motor base stops and moves upward to its original position. To deposit the
sample into a storage vial, the cutting device is moved to the storage area and is placed above an
empty vial. The motor base and the cutting tip move downwards until the bottom limit switch on
the main frame is activated. Then, the assembly stops and pressurized air expels the sample out
of the cutting tip.
68
Main base
Flexible shaft
holder
Linear
slide
Motor shaft
Limit
switches
Digital linear
actuator
Motor base
Cutting tip holder
Linear
slide
Cutting tip
Figure 3.41: The cutting device
The cutting device's specifications are as follows:
" Vertical motion of 1.5-inches
" Limit switches for sensing end positions, and sensing if cutting tip is touching gel tray
" Sample expulsion by pressurized air
* Different cutting tips for different band shapes
A vertical motion of 1.5-inches is sufficient since all the devices that the cutting tool is
interacting with are within this distance range. A voice coil was first selected as the vertical
actuator because of its simplicity and accuracy of motion. However, due to the high price of a
voice coil and controller system, this design was discarded. Instead, the proposed design uses a
Digital Linear Actuator (DLA) from Thomson Airpax Mechatronics as shown in the figure.
DLA is a modified rotary stepper motor. The rotor is internally threaded so that it mates to a
fixed lead screw shaft. When the rotor turns, the motor moves along the shaft. This design has
the advantage that it uses a minimum number of components since the shaft and motor are
69
included in one package. The selected DLA is a L92121-P12. It has a 1.875-inch travel distance
with a 0.002-inch resolution and a 26-ounce maximum force. The complete specification of the
L92100 motor and its driver is presented in Appendix D3. The lead screw shaft is connected to
the main base by a spring metal plate as shown in Figure 3.41. This provides flexibility to the
shaft to compensate for any misalignment between the shaft and the rotor.
Two linear slides from IKO, model BSP 10-45 SL and BSP 10-25 SL, are used to hold
respectively the motor base and the cutting tip base. The BSP series slide is a lightweight and
compact stainless steel rolling glide. It has an accuracy of 0.004mm in the vertical direction and
0.008mm in the lateral direction.
Three limit switches are necessary for the proper operation of the cutting device. Two limit
switches on the main base initialize and sense the maximum positions of the motor base. The
third limit switch, which is not shown in the figure, is attached to the motor base and sense when
the cutting tip base is touching the gel tray. This limit switch adds two important features to the
designed cutting tool. No calibration of the vertical position of the cutting tip relative to the gel
tray is needed since the cutting tip will automatically stop once it has reached the gel tray
surface. Furthermore, this is also a safety feature because if the cutting tip hits an unexpected
obstacle while moving downward, it will stop automatically.
The force required to move the
cutting tip holder relative to the motor base is determined by a spring located between the two
frames as seen in Figure 3.41. The spring has to be stiff enough so that the cutting tip is able to
cut the gel, but not too stiff to avoid scratching the gel tray.
The expulsion of the sample from the cutting tip is done using pressurized air. A flexible
tube is connected to the back of the cutting tip holder and a small hole passes the air through the
cutting tip and its holder to the inside of the tip.
The cutting tip shown in Figure 3.42 is magnetically attached to its holder.
A precise
alignment of the cutting tip on its holder is made possible by two guide pins on the holder that
fits in the position holes of the cutting tip. In order to accommodate the different sizes and
shapes of the bands to be excised, cutting tips with different blade sizes are used.
70
Pressurized
air hole
Magnet
Positioning
holes
Stainless steel
blades
Figure 3.42: The cutting tip
The tip of the cutter is made of extra sharp stainless steel blades of 0.009-inch thickness. The
blade cover was removed and the blades were cut into strips to form the walls of the tip. The
strips were then glued together using epoxy. These tips were made only to prove the concept.
For the final design, a different process is needed to make the rectangular tips. This issue is
discussed in more detail in the next section.
3.5.5
Results and Recommendations
An experiment was conducted to check the ability of the cutter to excise and expulse a band from
an electrophoresis gel. For this experiment, the cutting tip had an internal dimension of 1mm x
5mm. Two agarose gels of 1mm and 6mm respectively were tested to check the influence of gel
thickness on the cutting process. To expel the band from the cutting tip, a syringe was connected
to the air tube. In this test, the cutter was moved manually onto the gel. The results of this
experiment are as follows.
*
The cutter was able to cut a band for both the 1mm and the 6mm gel. However, it
was observed that when the cutter came in contact with the gel surface, the gel
surface compressed first for a short vertical distance vertical of around 0.1-0.2mm,
and then the blade cut through the gel. This phenomenon did not affect either the
shape or integrity of the gel and the extracted band. If necessary, this behavior could
be avoided by inducing small ultrasonic vibration in the blade with a piezoelectric
actuator.
The ultrasonic vibrations would facilitate the cutting process and also
prevent small unwanted-particles to stick to the cutting tip.
71
* The cutter extracted the 6mm thick bands, whereas the 1mm bands were not always
pickup from the gel. An examination of the razor blade can provide some insight into
this observation. The razor blade is sharpened on both sizes as seen in Figure 3.43a.
The height of the edge is 1mm, which is the same height as the gel. Therefore, the
band is being pushed outward by the walls. To eliminate this problem, a cutter that is
only sharpened on the outside is necessary as shown in Figure 3.43b.
Such a
component can be made - either of plastic or metal - by the extrusion of a hollow
rectangular bar and the sharpening of the outside edges.
Cutting blade
Cutting blade
Desired band
Desired band
Imm
mm
5mm
5mm
b) Improved design
a) Current design
Figure 3.43: Cutting tips cross section
*
To expel the sample out of the cutting tip, the syringe was used to send air inside the
tip. It was observed that the bands came out, but remained stuck on one side as
shown in Figure 3.44. The location of the air hole causes the observed phenomena.
The orifice was placed on the side of the cutter and therefore the air only pushed one
side of the band.
Air hole
M
Positioning
pin hole
n
Cutting tip base
Cutting tip
Desired band
Figure 3.44: Band expulsion test result
72
To improve this design, the air path needs to be changed so that the air jet is pointed
towards the center of the band. The location of the air orifice on the top cannot be
changed because the magnet has to be close to the center to ensure a good connection
between the cutting tip and its holder. The two possible modifications to the orifice
location are shown in Figure 3.45. The first one is shown in Figure 3.45a, and
consists of inclining the air orifice so that the air jet is pointing toward the center of
the cutting tip. In the second modification, as seen in Figure 3.45b, the cutting tip is
moved to the side so that it is centered on the air orifice. The orifice diameter should
also be larger to increase the volumetric flow rate of the air entering the cutting tip.
Air hole
Air hole
Cutting tip base
A-
Cutting tip base
Cutting tip
Cutting tip
Desired band
Desired band
a) Inclined air orifice
b) Off-center cutting tip
Figure 3.45: Possible modifications of the cutting tip expulsion system
*
It was also observed that, when pressurized, some air leaked from two places in the
device: the sharp end of the cutting tip and the interface between the cutting tip and
its holder. The blade being sharpened on both sides causes the leak at the cutting tip
end. This problem will be resolved by using the new type of cutting presented in
Figure 3.43b. Improving the joint design between the cutting tip base and its holder
can minimize the leak at the interface. This could be accomplished with a gasket.
In conclusion, the mechanical stamping approach is effective in excising a band out of an
electrophoresis gel. Some of the proposed changes need to be reviewed and implemented to the
current design to improve its effectiveness and reliability.
73
3.6 Design of the Cutter Transportation System
Once a desired band is excised from the gel, the cutter system needs to be moved to the storage
area in order for the band to be deposited there. An automated transportation system for the
cutting tool is presented in this section.
3.6.1
Requirements
The purpose of the transportation system is to quickly move the cutter between the different
stations. The following are its requirements:
" Able to carry the cutting tool between stations.
*
Capable of reaching a maximum speed of 6-inch per second.
" Having a linear resolution of 0.5-millimeter.
The only process in the entire machine that need high precision placement is the band
excision process. The XY stage provides a positional accuracy of 0.075mm, which is ample for
that process. Therefore, the transportation system can have a coarser precision. An accuracy of
0.5mm is selected since that will allow a repeatable placement of the cutter at each station.
Having a lower precision has the advantages of being able to reach higher speeds.
3.6.2
Possible Designs
There are three major designs for transportation systems, pulley and belt, lead screw, and rack
and pinion. Figure 3.46 illustrates them.
a)
c)
b)
Figure 3.46: Transportation systems: a) pulley-belt, b) lead screw, c) rack-pinion
The pulley and belt system is used in application where it is needed to move a light object
rapidly between two points. See Figure 3.46a. It is an efficient way of moving an object for
short travel distance but not for long travel distance.
A long belt results in an increase in
backlash, and a decrease in positioning accuracy due to the belt flexibility. On the other hand,
74
the lead screw system, shown in Figure 3.46b, can have high resolution, but then moves slowly.
It is also a more complex system since the lead screw has to be properly supported. The rack and
pinion system is shown in Figure 3.46c.
Its performance lies in between the two previous
systems. It has a good balance of traveling speed and positioning accuracy that makes it the
ideal choice for the proposed automated system.
3.6.3
Design of the Transportation System
The final design of the transportation system is shown in Figure 3.47. The assembly and the
detail drawings are shown in Appendix E2. The transportation system is composed of four
subassemblies: frame, sliding mechanism, rack and pinion, motor and its holder.
-
Frame
Slide
Rail
Stepper
motor
Racks
Cutter
holder plate
Pinion gear
Figure 3.47: The cutter transportation system
The cutting device transportation system specifications are as follow:
*
Maximum speed of 150 mm/s at 300 pulses per second.
* Linear resolution of 0.5 millimeter.
*
Starting acceleration of 150 mm/s2 .
" Linear distance range of 850 millimeter.
75
Two types of sliding mechanism can be used in our automated system, a shaft-bushing
system, and a rail bearing system. The rail system was chosen because it is a smaller and a more
easily implemented system.
An AccuGlide miniature linear ball bearing from Thomson
Industries, Inc was selected. It provides a smooth, quiet linear motion and is designed for standalone application.
The rack, gear, and motor were selected together because they have interdependent
characteristics.
The pitch for the rack and pinion system needs to be as large as possible to
obtain a smooth motion. However, as the pitch increases, the lesser the load that can be applied
to the teeth of the rack and gear. A 48 pitch was selected because it has a good balance of gear
tooth strength and linear pitch distance. The linear pitch is 1.7mm and the gear has a tooth
strength of 122N. The diameter of the pinion gear determines the linear resolution and the speed
of the system. No gear trains are used between the motor shaft and the pinion because that
would result in a large backlash and a lower efficiency. Instead, the pinion gear is directly
connected to the motor shaft. The stepper motor needs to have small step angle to achieve the
desired resolution. A 4SQ-120BA34S stepper motor for Thomson airpax mechatronics was
purchased. It provides 37mN-mm of torque at 300-pulse-per-second, and has 200-steps-perrevolution. An anti-backlash pinion gear with a pitch diameter of 1.25-inch from Berg, Inc was
then purchased to achieve the desired linear resolution of 0.5mm and a speed of 150mm/s. The
derivation for the required stepper motor torque and the system specifications are shown
Appendix El.
The motor driver and electronics were designed and are shown in Appendix A4. The motor
is controlled through a program developed in C++ builder 4 by a project member. An example
of the interface screen in shown in Appendix H1. The program controls the direction and the
linear speed of the cutter transportation assembly. Each pulse sent through the D/A card to the
motor driver results in a step of 0.5mm. For proper operation of the motor, a ramping scheme
was implemented in the software for the pulse rate. The assembly accelerates to cruising speed
in about 1.5-second. The pulse rate starts at 60-Hz, and then increases linearly until the desired
final speed is reached.
The same ramping scheme is also implemented for the deceleration
section of the motion. A magnetic reed switch is used to calibrate the system.
76
3.6.4
Results and Recommendations
The transportation system was built and tested.
Two tests were performed to check the
repeatability of the transportation system. The first test consisted in calibrating the stage to the
home position and then sending it 600mm away. This test was performed 20 times. The stage
repeatedly calibrated itself correctly for all the runs. For the position test, 1200 pulses were sent
to the driver, and the final location was measured. Out of the 20 runs, 19 times the stage moved
exactly 60cm, and once it moved 59.5cm. Therefore, the motor missed one step out of twelve
thousands once. The second test consisted of making the transportation system perform a series
of movement and checking its position at every location. This test was also successful and it is
concluded that transportation system is working properly. In the final design, a position encoder
needs to be added to the system to ensure an accurate positioning of the carriage along the track.
During the system assembly, it was observed that the alignment between the pinion gear and
the racks is critical for a smooth operation. The system was designed with four racks of 9-inch
to allow the user to easily increase or reduce the traveling range of the transportation assembly.
However, this greatly complicated the racks alignment process. In the next generation, a onepiece rack should be used instead.
77
3.7 Cutting Tips Changing Station
Once a desired band is excised from the gel and is deposited in the storage area, the used cutting
tip needs to be exchanged with a clean one before the next excision. A cutting tip changing
station is presented in this section.
3.7.1
Requirements
The purpose of the cutting tips changing station is to grab the cutting tip from the cutting tool
and store it. The main requirements are to:
" Remove the cutting tip from the cutting tool.
*
Store at least four cutting tips.
"
Store different cutting tip sizes.
The station needs to store different cutting tip sizes so that a wide range of band shapes can
be excised. Two cutting tips of each size need to be stored so that if the same size is necessary in
successive cuts, one tip is being cleaned while the other is used.
3.7.2
Design of the Tool Changing Station
Two possible mechanisms to remove the cutting tips from the cutter and to store them have been
developed: an electromagnetic design and a mechanical clamp.
The electromagnetic design consists of using a strong electromagnet to attract and hold the
cutting tip onto the changing station. The electromagnet needs to be placed below the platform
that holds the cutting tips. The tool station magnet power has to be large enough to overcome
the attractive force between the cutting tip magnet and the cutting tip holder. This design has the
advantage of requiring only a few components. However, holding the cutting tip accurately at its
stored location might be difficult to achieve when the magnet is not energized. A self-centering
platform needs to be used.
The mechanical clamp design consists of physically holding the cutting tip by using a clamp
as shown in Figure 3.48. This design has the advantage of ensuring reliable storage of the
cutting tip and therefore will be used in the design of the tool changing station.
The final design of the tool storage station is shown in Figure 3.48. The assembly and the
detail drawings are shown in Appendices F1.
The tool station is composed of four
subassemblies: frame, sliding mechanism, solenoid actuator, and locking mechanism.
78
Front
Back
spring
Shaft holder
Transparent
base plate
spring
Cutting tip
centering
grooves
Solenoid
Sliding
shaft
Locking
setscrew
Hole for
cutting blade
Delrin
sliding arm
Figure 3.48: The cutting tip storage station
The operation of the tool changing station is as follows. To drop off a cutting tip, the
solenoid is first energized to open the clamping mechanism. Then, the cutting device lowers the
cutting tip to be cleaned in an open slot. Once in position, the solenoid is deactivated, thus
releasing the sliding arm and allowing it to clamp on the cutting tip. Finally, the cutting device
moves back up leaving the cutting tip behind. The sequence to pick-up a cutting tip starts with
the cutting device moving down on the desired tool. Then, the sliding arm moves back to release
the cutting tip. Finally, the excising device moves upward with the tip attached to its holder.
The cutting tool changing station specifications are as follows:
" Holds five cutting tips.
*
Actuated by a pull-type, 12-Oz force solenoid.
*
Soft closure motion to ensure a reliable tool grabbing.
" Cutting tips are secured in placed in the default/empowered position.
79
In the final design, the tool changing station is incorporated with the cutting tip cleaning
station. Therefore, the blades of the cutting tip need to protrude under the changing station. For
this reason, the base frame is made out of a sheet of clear plexiglass. The transparency of the
base will allow the user to see the cleaning process from above. As seen in Figure 3.48, five
tools can be stored at the same time. The sliding arm is holding them in place. The arm is made
out of delrin material. Delrin is ideal for our application because no bushings are necessary if
used to slide on stainless steel and it provides some viscous damping that will slow down the arm
motion. When the solenoid is not energized the sliding arm position is controlled by two sets of
springs. The back springs push the sliding arm onto the cutting tips to hold them at their
respective position and the front springs act as kinematic dampers to slow down the locking
motion. The shaft holder position can be changed to adjust the force exerted on the cutting tips
by the sliding arms. The solenoid can also be moved to vary the sliding arm traveling distance.
As seen in the figure, setscrews with plastic heads are mounted on the sliding arm. Their
purpose is to securely hold the cutting tip during its removal from the cutting device. There are
also centering grooves both on the base plate and on the fixed part of the clamping mechanism.
They ensure that the cutting tip is correctly positioned during the clamping process.
The
computer controls the solenoid. A transistor type circuit is used for the power switching. A
schematic of the electronics is shown in Appendix A4.
3.7.3
Results and Recommendations
The tool changing station was built and tested. During experimental tests, the station repeatedly
centered and locked the cutting tips in place. The input voltage of the solenoid for opening and
closing the clamping mechanism is shown Figure 3.49. The solenoid can open and release the
clamp mechanism quickly. However, this would cause the sliding mechanism to collide with the
cutting tip when closing and the solenoid frame when opening. Springs are used to slow down
the motion. Consequently, the sliding mechanism impacts are significantly reduced. In the final
the clamps opens in 1 second, and closes in around 3seconds.
The closing is intentionally
slower. It smoothes the motion ensuring that the cutting tips are properly locked in place.
80
1
-
-
1
.~
~
So enoid
de-e nergize
d n up ddse
mp opEns
Figure 3.49: Tool changing station solenoid input voltage versus time
The designed tool changing station can hold five cutting tips. That number can be changed
by reconfiguring the holes' position. As seen in Figure 3.50, a multiple layer or even a circular
pattern could be easily implemented.
Figure 3.50: Other designs to increase number of stored cutting tips
81
3.8 Cutting Tip Cleaning Station
The cutting tips need to be cleaned between excisions.
They are removed from the cutting
device and stored at the tool storage station. The blades of the cutting tip extrude by 9mm under
the base plate of the tool station. This allows the blades to be cleaned by the cleaning station. A
cutting tip cleaning station is presented in this section.
3.8.1
Requirements
The requirements of the cleaning station are simple, yet very difficult to achieve. They are:
" To completely remove contaminants from the cutting tip blades,
" To leave the cutting tip free of cleaning chemicals,
*
To have a short cleaning cycle.
Each band in the gel contains different molecules. Therefore, it is critical to avoid sample
cross contamination between excision of different bands.
Since the cutting tips are used
repeatedly, they need to be thoroughly cleaned. The cleaning process has to remove all traces of
gel from the cutting blade as well as removing any chemicals such as the gel buffer. To reduce
the excision cycle time, the cleaning process should be as short as possible.
3.8.2
Possible Cleaning Methods
Five different cleaning methods have been reviewed for the cleaning station: external spray,
spray in cutting tool, liquid jet, ultrasonic wave, vibrating tool. Figure 3.51 illustrates them.
The spray system is shown in Figure 3.51a. It consists of three nozzles to disperse the
cleaning fluid on the cutting blade. The contaminated fluid is then collected at the drain and is
recycled by passing it through a filter for reuse. In this system, the nozzles are located in the
cleaning station. They could also be placed in the cutting tip itself as shown in Figure 3.51b.
Two nozzles spray on the outside of the tip, and one sprays in the inside. The advantage of this
method is that the inside nozzle can more effectively clean the inside of the cutting tip than the
external nozzle of the previous design. This is because with the internal nozzle, the liquid enters
from the top and exits at the bottom, whereas with the external nozzle, the liquid enters and exits
at the bottom. However, placing three nozzles on the cutting tip is difficult due to its small size.
An alternative approach would be to combine the two methods by using one nozzle on the
cutting tip spraying the inside and two other external nozzles for the outside of the tip.
82
Cutting tip
liqui~dt
nozzle
Vibrates using
a)
stepper
Filter
Pump
4-1
b)
+
Piezoe ectn
e)
actuator
Figure 3.51: Different cutting tip cleaning methods
The next three methods involve the immersion of the tip in the cleaning fluid. The liquid jet
system is shown in Figure 3.51c. A nozzle creates a turbulent flow that cleans the blade. This
method is simple to implement. However, the jet power might not be sufficient to fully clean the
cutting tip. The liquid jet coming out of the nozzle is dispersed quickly because it is passing
through a liquid with the same viscosity. An alternative would be to create a turbulent flow
around the cutting blade by vibrating the cutting tip itself as shown in Figure 3.5 1d. The cutter's
stepper motor could be used for this purpose. For this method to be effective, the cutter needs to
be vibrated both horizontally to clean the outside of the tip and vertically to clean the inside
walls. The final method, ultrasonic cleaning, is shown in Figure 3.51e. Ultrasonic cleaning can
remove contaminants such as buffing compounds, dried blood, oil, greases, and surface debris.
This method can be used on a variety of materials such as metals, glass, plastic, and ceramics. It
can penetrate and thoroughly clean microscopic crevices.
Of all the presented methods,
ultrasonic cleaning is the most appropriate method for this tool because of its cleaning efficiency
and effectiveness. Therefore it is the selected method in the design of the cleaning station.
83
3.8.3
Design of the Cleaning Station
An understanding of the ultrasonic cleaning process is necessary to properly implement such a
system.
Therefore a brief explanation is now presented.
For more details, please consult
appropriate literature such as References [12] and [13].
A typical ultrasonic cleaner is shown in Figure 3.52. A piezo-electric transducer is attached
to the bottom of the tank and is used to send high-frequency sound waves in the 20-40 kHz range
through the liquid. The pressure fluctuation causes micro-sized vapor bubbles to form in the
liquid. Since there is insufficient energy in the liquid to sustain the vapor state of the bubbles,
they collapse violently. This phenomenon is called "cavitation". When collapsing, the pressure
in the micro-bubble is on the order of 500atm and the temperature ranges from 50000 C to
10,000*C. When the micro-bubble collapses, it creates a jet of about one-tenth the bubble size
with speeds of up to 1 lOm/s. When this happens next to a contaminated surface, the jet removes
the unwanted molecules from the surface.
Because of the small jet size and high energy
released, the cleaning solution can easily penetrate and clean small crevices and tightly spaced
parts. [12]
Three main parameters affect the effectiveness of the ultrasonic cleaning process: the
cleaning chemical, solution temperature, and solution degassing
The selection of the cleaning chemical is essential to the cleaning process. The chemical has
to be capable of removing the contaminants and also be compatible with the material being
cleaned. Most cleaning chemicals can be used with ultrasonic cleaning.
The temperature of the solution is an important factor in optimizing the cavitation process.
In general, a rise in temperature will increase the cavitation intensity, and consequently will
result in better cleaning. However, the temperature should always be below the boiling point of
the solution because the cavitation intensity will be significantly reduced as the solution starts
boiling. For instance, the cavitation effect is maximized at around 71C for a water solution.
The cleaning solution should be degassed to increase the cavitation intensity.
When a
cavitation bubble forms, any dissolved gas in the solution will diffuse into the bubble. As the
bubble collapses, these gases will create a visible bubble since they cannot diffuse fast enough
into the liquid.
This effect reduces the cavitation intensity.
However, as the sonification
continues, these bubbles of gas will coalesce and rise out of the liquid thus degassing the
84
solution. Therefore, the solution should be degassed prior to the cleaning process by operating
the ultrasonic cleaner until no bubbles are seen rising to the surface of the solution. [13]
In summary, ultrasonic energy can effectively and efficiently clean the cutting tips. A model
UC-1 ultrasonic cleaner from Electrowave Corporation was purchased and is shown in Figure
3.53. This unit was selected because it has the smallest tank capacity found. It has a capacity of
270-mL and a peak power of 100-Watts.
Figure 3.52: The UC-1 ultrasonic cleaner
3.8.4
Results and Recommendations
The ultrasonic cleaner was tested to check its cleaning effectiveness. The goal of this experiment
was to mimic its future application. Two different solutions were tested: distilled water and a
general-purpose aquasonic cleaning solution from VWR Scientific. The cleaning solution was
selected because it is compatible with a variety of substrates, especially stainless steel and
aluminum, and because it is environmentally friendly. The same stainless steel razor blades that
are used to make the cutting tip are used for the substrate. The project sponsor suggested using
writing inks as contaminant samples since they can be easily visualized.
Five different
contaminants were tested on the blades:
*
Black ink from a ball pen
*
Black ink from a permanent marker
*
Red ink from a dry erase marker
*
Graphite particles from a #2 pencil
*
General purpose oil
The first three were selected to test different type of ink. Graphite left by a pencil mark is
made up of many small particles, which provides a fourth test contaminant. Oil is used as a test
85
contaminants to check the ability of the system to remove external contaminants such as
fingerprints.
Two identical groups of five blades were prepared: one for the solvent test and one for the
water test. Four spots of the same contaminant were deposited on each blade in a group. The
blades were then allowed to dry for 3 hours. The five blades were mounted on a plastic support
and placed in the tank of the ultrasonic cleaner with the correct amount of cleaning solution as
shown in Figure 3.52.
The blades cleaned with the solvent and water solutions for each contaminants are shown at
various time throughout the cleaning process in Table 3.5 and 3.6
Solvent Cleaning Solution
Water Solution
Before
Before
-
Black ink
from a ball pen
------1 minute
1 minute
3 minutes
6 minutes
3.5 minutes
i
Black ink from a
permanent marker
i
Before
Before
1 minute
1 minutes
3 minutes
6 minutes
S
Table 3.5: Results of the ultrasonic test for different solution
86
77-77
e
Solvent Cleaning Solution
Water Solution
Before
Refore
Red ink from
a dry erase marker
-mnt
I minute
1 minutes
Before
Betore
Graphite particles
from a #2 pencil
minute
1 minutes
3 minutes
6 minutes
I
I
i
Refnre
Hetore
General
purpose oil
<30 seconds
1 minutes
%-I
Table 3.6: Results of the ultrasonic test for different solution
87
111111L&L% k3
Two observations can be made from this experiment.
First, the solvent solution unlike the water solution was able to remove all the contaminants
from the blades.
contaminant.
The water did however have a limited effect on the blade with the oil
This confirms the importance of the solution in the cleaning process.
The
cavitation effect can be pictured as a microscopic scrubbing effect. Its purpose is to accelerate
the cleaning cycle and to increase the cleaning capabilities.
Second, in the solvent solution experiment, the time required to clean the blades is dependent
on the type of contaminant used. The oil was removed under 30 seconds. The dry erase ink and
the graphite were removed in about 1 minute. The permanent ink and the ball-point pen ink took
about 3 minutes to be extracted from the blade surface.
A second experiment was developed and performed to check if the cleaning time could be
reduced further. It consists of not using the plastic blade mounts and instead placing the blade
directly on the bottom of the tank so that it touches the tank's vibrating surface. The results are
shown in Table 3.7.
Before sonification.
Lines are from left to right:
ball pen, permanent marker,
dry erase marker, pencil, and oil
After 30 seconds of sonification
After 2 minutes of sonification
Table 3.7: Test result for blade cleaned while touching the bottom of the tank
As seen in Table 3.7, the cleaning cycle time in this set-up is about half as long as the set up
using the mounts.
One possible reason for this observation is that by touching the bottom
surface, the blade vibrated thus accelerating the cleaning process.
88
In conclusion, the ultrasonic cleaner was successful in cleaning the cutting blades. However,
a chemical based solution needs to be used to effectively remove chemical contaminants from
the cutting tip. This means that the cutting tip needs to be rinsed after being cleaned to remove
the cleaning solvent.
Water can be used for this purpose.
To accommodate for this new
requirement, two beakers need to be placed in the tank of the ultrasonic cleaner, one filled with a
solvent cleaning solution, and the other with a rinsing solution. Figure 3.53 shows such a set-up.
Figure 3.53: Ultrasonic cleaner assembly using one beaker for cleaning and one for rinsing
89
3.9 Temperature Controlled Sample Storage Station
Once a band is excised from the gel, it needs to be stored until it is needed for further analysis.
A temperature controlled storage station for the excised samples is presented in this section.
3.9.1
Requirements
The main requirements of the sample storage station are:
" Storage for at least 20 different samples,
" Temperature controlled from 4-900 C,
*
Minimal possible size.
The storage station has to be able to store different bands in individual container since the
user might be interested in excising several bands from the gel. Depending on the type of postanalysis desired, the sample needs to be stored at a specific temperature.
3.9.2
Design of the Temperature Controlled Storage Station
The operating temperature of the storage station ranges from below to above the ambient
temperature.
The simplest and most efficient method of achieving this consists of using
Thermoelectric Coolers (TEC). TEC or Peltier coolers are solid-state heat pumps. A schematic
diagram is shown in Figure 3.54. Applying a DC voltage across its leads will cause the heat to
move from one side of the TEC to the other. Changing the polarity will result in the heat moving
in the opposite direction. Consequently, a TEC can be used for both cooling and heating.
Cooled Object
Electrical connector
N
P
N
P
Heat Sink
DC Power Supply
Figure 3.54: Schematic of a thermoelectric cooler
90
Electrical Insulator
Good heat conductor
A typical thermoelectric module consists of a series of P and N doped bismuth-telluride
semiconductor material sandwiched between two ceramic plates. The N type material has an
excess of electrons, and the P type material has a deficit of electrons. A pair of each makes a
couple and there are between one and a few hundred couples in a TEC. When an electron moves
from a P to an N type material on the cold side, it jumps to a higher energy state thus absorbing
some thermal energy. On the hot side, the electron moves from an N to a P type material,
dropping to a lower energy state. This results in a release of thermal energy to the heat sink.
The final design of the temperature controlled storage station is shown in Figure 3.55. The
assembly and technical information for each component are presented in Appendix G2 and G3.
The station is composed of three subassemblies: the thermoelectric coolers system, the storage
enclosure, and the power supply.
Figure 3.55: The temperature-controlled sample storage station
Two 127-couple, 8.5-amp thermoelectric cooler modules from Ferrotec Inc. were purchased.
A model 5C7-350A temperature controller from Oven Industries, Inc. is used to control both
modules. It provides proportional and integral control of the temperature within a range of -20'C
to 100*C.
Two CPU cooling fans with heat sinks are used to dissipate the heat from the
thermoelectric coolers. The hot side of the thermoelectric module is bonded to the heat sink with
91
a thermally conductive epoxy. The cold side is coated with thermal conductive grease and is
clamped to the stainless steel pan. The DC power supply is from Power-one Inc. and can output
up to 28-volts at 6-amps.
The enclosure is composed of a stainless steel pan insulated on the outside. A model TS67178 thermistor from Oven Industries Inc. was bonded to the side of the pan. It provides the
temperature feedback for the controller. Expanding polyurethane foam was applied around the
sides and part of the bottom of the pan to insulate it. An aluminum rack that can hold up to 24
1.5-2.2 ml micro-centrifuge tubes is mounted inside the pan.
The calculations for the storage system parameters are shown in Appendix G1.
3.9.3
Results and Recommendations
The temperature controlled storage station was tested under various conditions to check its
effectiveness. Only one of the two cooling modules was used for these tests.
The first test consisted of measuring the system response to a step input. While at room
temperature, the temperature control knob was set to its minimum.
The temperature
measurements were made at the bottom of the pan directly above the module location. Once the
system stabilized, the knob was set to its maximum. The results are shown in Figure 3.56.
Peltier cooler cold side surface temperature
Power supply at 12 Volts and 5 Am ps
30
27.5
-+-
--
Cooling
Fbw er off
25
22.5
.
20
L 17.5
15
12.5
E
10
7.5
5
2.5
UI
0
20
60
40
80
100
120
Time (s)
Figure 3.56: Temperature on the thermoelectric cold side during power-up and power-down
As seen in Figure 3.56, the response has the characteristic of a second order system. The rise
time was approximately 10 seconds and the settling time was around 50 seconds for both the
92
powering-up and powering-down of the module. The oscillations in the temperature are caused
because the module was only in contact with ambient air and thus it had a small heat load. When
powering-down, some of the heat from the heat sink is transferred by conduction to the cold side
of the module. This results in a peak temperature that is above the room temperature, as seen in
the powering-down response.
It should be noted that during the test, the minimum temperature the system achieved for a
prolonged period of time was 0.5"C. However, theoretically, the module should be able to reach
-20*C. This discrepancy is caused by the inability of the system to dissipate the heat generated
by the module fast enough. There are two solutions to this problem. First, replacing the current
heat sink and fan by higher performance components would increase the heat dissipation. This
would result in a lower temperature at the hot side of the module and therefore a lower
temperature at the cold side. Second, two modules could be stacked on top of each other so that
one of the module is used to cool the other one. In this set up, the modules would be connected
thermally in series and electrically in parallel.
The second test consisted of measuring the temperature at a height of 10mm from the bottom
of the pan. The test was performed with three different mediums in the pan: still air, 20-mm of
still water, and 20-mm of continuously mixed water. The results are shown in Figure 3.57.
Cooling test for different heat transfer materials
Temperature measurements at 10mm from bottom
+ Still water (20mm heigth)
25
-0- Mixed water (20 mm heigth)
24
-*-Still air
2322
~21
1716-
15
0
2.5
5
7.5 Time (min)
12.5
15
17.5
20
Figure 3.57: Temperature at a height of 10-mm with different heat transfer materials
93
As seen in Figure 3.57, the temperature of the mixed water medium dropped the fastest,
followed by the still water medium and finally by the still air medium. This result is to be
expected since water has a higher thermal conductivity than air. Mixing the medium also
accelerates the cooling because it increases the heat transfer coefficient. However, for all three
set-ups the rate of cooling is relatively slow. The mixed-water cooled from 22*C to 15.7 0C in 12
minutes. This is not fast enough for the desired application. One possible approach to improve
the performance is to look at the thermal properties of the materials used.
A few selected
material properties are shown in Table 3.8.
Material
Density
gm/cm3
Air
Aluminum
Copper
Gold
Stainless steel
0.0012
2.71
8.96
19.32
8.01
Water
1
Thermal
conductivity
Watts/m-"C
0.026
204
386
310
13.8
0.24
0.215
0.092
0.03
0.11
.61
1
Specific heat
cal/gm-0 C
Thermal expansion
coefficient
Cm/cm/'C
22.5
16.7
14.2
17.1
Table 3.8: Selected material property at 21'C
For the temperature-controlled station, the heat transfer between the module and the stored
sample needs to be maximized. Therefore the medium used to transfer the energy has to have a
high thermal conductivity to increase the heat transfer. A low specific heat is also important to
reduce the time required to change the temperature of the medium. As seen in Table 3.8, air is a
thermal insulator and water needs a lot of energy to change its temperature. Aluminum is a
better choice since its thermal conductivity is 330 times greater than water and its specific heat is
one-fifth that of water.
A third test was run to check this hypothesis. An aluminum block was placed in the pan, and
the temperature was measured at a 10mm height from the bottom. The system was first turned
on to cool the block, then, once the temperature had settled, the module power was turned off.
The results are shown in Figure 3.58. A heating test was also performed. The thermocouple
power supply was turned on and the temperature was measured at the same location used for the
cooling test. Once the system settled, the power supply was turned off and the temperature was
recorded for 20 minutes. The results are shown in Figure 3.59.
94
Cooling test with an aluminum block
Temperature measurements at 10mm from bottom
24
22
20
18
16
14
S12
10
8
E
cooling
__ power off
-
6
4
2
0
0
1
2
3
4
Time (min)
5
6
7
8
Figure 3.58: Cooling test, temperature at a height of 10-mm with an aluminum block
Heating test with the aliminum block
Temperature measurements at 10mm from bottom
120
- heating
-a- power off
100
80
60
40
0
0
2
4
6
8
10
12
Time (min)
14
16
18
Figure 3.59: Heating test, temperature at a height of 10-mm with an aluminum block
95
20
As seen in Figure 3.58, the system performance significantly improved with the use of the
aluminum block as the heat transfer medium. The set-up with mixed water reached 16 0C in 12
minutes, whereas this set up reached 4*C in 4 minutes. The settling temperature was 3*C, which
is close to the 2*C temperature at the surface of the pan.
For the heating test, the temperature reached 100 0C. The rise time and settling time for heating
the block were 2.5 minutes and 4.5 minutes respectively. These results are similar to the ones
observed for the cooling test.
The results of these tests suggest that an aluminum block needs to be used to hold the storage
tubes since it significantly improves the performance of the system.
In conclusion, the first prototype of the temperature controlled storage station is working well.
In future work, the heat sink performance must be improved by implementing one of the two
aforementioned solutions. The tank is not needed and should be replaced with an aluminum
block that is machined to fit the micro-centrifuge tubes.
96
Chapter 4
Conclusion
The first part of this thesis presented a stain free detection system that can be used for the
purification of a variety of biologically active macromolecules. The method is based on direct
UV absorption and takes advantage of the latest development in CCD technology to obtain high
detection sensitivity.
The second part of the thesis presented six main components of the robotic system: a variable
ultraviolet light source, a gel cutting device, a cutter transportation system, a cutting tip changing
station, a cutting tip cleaning station, and a temperature controlled sample storage station.
The variable light source was developed to allow for a precise selection of the target
molecules to be visualized. It outputs a uniform, collimated beam with a 32mm cross-sectional
diameter and a 15nm bandwidth over the 200-400nm light spectrum range. The light source can
be used either as a monochromatic light source to visualize the location of a specific molecule, or
as a scanning instrument to obtain the absorption spectrum of a bio-molecule. The light source
meets the functional requirements, however its performance could be improved. The use of a
larger grating could increase the throughput. The bandwidth of the light output beam could be
reduced by either using baffles or by replacing the lamp with a projecting type lamp. Finally, the
effects of varying the slit height and width on the output beam characteristics need to be
investigated.
The mechanical cutting device can excise bands of various shapes and sizes from an
electrophoresis gel. It has a 1.5 inch vertical travel and automatically senses when a cut has been
made. The sample is expelled from the cutting tip using pressurized air. It is recommended to
change the cutting tip blades by ones that are sharpened only on the outside to ensure the pick-up
of the band from the gel. The sample expulsion needs to be improved by changing the location
of the pressurized air orifice.
The transportation system was developed to move the cutting device between the different
stations. The system has a 0.5mm linear resolution and an 850mm traveling distance. The
97
moving assembly can travel at speeds of up to 150mm/s. In the final design, a one-piece rack
should be used to facilitate assembly. A position encoder should be added to the system to
ensure an accurate positioning of the carriage.
The cutting tip changing station removes the excision tip from the cutting device and stores
up to five tips. In the next prototype, the base platform needs to be redesigned to fit on top of the
cleaning station.
The cleaning station uses ultrasonic energy to effectively and efficiently cleans the cutting
tips. The cleaning is done in two stages. First, a solvent solution removes contaminants from the
blades. Then, a rinsing solution is used to remove the solvent from the blades. In future work, a
filtering system could be added to the station to recycle the cleaning solutions.
The temperature-controlled station uses solid-state air-cooled thermoelectric devices. It can
store 24 micro-centrifuge tubes and has a temperature range of 4*C to 100*C.
A lower
temperature can be achieved by using a higher performance heat dissipation system.
In conclusion, the proposed system has been successfully designed and implemented.
However, it is the first prototype.
Additional features need to be incorporated prior to
commercialization.
98
Bibliography
[1] "Gel Electrophoresis and photography AB-1000-02",
http://www.uvp.com/html/bulletins.html
[2] Kuhn and Hoffstetter-Kuhn , CapillaryElectrophoresis:Principleand Practice,
Springer-Verlag Berlin (1993)
[3] "Detection in the sheath flow cuvette",
http://hobbes.chem.ualberta.ca/~chris/ceover/cesheath.html
[5] Goeller, J.P and Sherry. "Ultraviolet photography of paper chromatograms in the study
of nucleic acids". Proc. Soc.Exp.Biol. Med. 74:381-382 (1950)
[6] Orson, F.M. "Photodocumentation of UV Shadowing with DNA Gels". Benchmarks
Vol. 16, No. 4, 592-595 (1994)
[7] Thurston, S.J and Saffer, J,D. "Ultraviolet Shadowing Nucleic Acids on Nylon membranes".
Analytical Biochemistry 178,41-42 (1989)
[8] Hendry, P., Hannan, "G. Detection and Quantitation of Unlabeled Nucleic Acids in
Polyacrylamide Gels", Biotechniques 20:258-264 (February 1996)
[9] Mahon, A.R., et al, "A CCD-based system for the detection of DNA in electrophoresis gels
by UV absorption", Phys. Med. Biol., 44, 1529-1541 (1999)
[10] Hecht, E. Optics, Third edition, Addison-Wesley
[11] Richardson Grating Laboratory, Diffraction GratingHandbook, Fourth edition, NY
[12] ASM International, "Ultrasonic Cleaning", ASM Handbook, Vol. 5, Surface Engineering,
p44-47, ASM International, Material Park, OH 44073-0002 (copyright 1994)
[13] Fuchs, John F., "Ultrasonic Cleaning: Fundamental Theory and Application"
http://www.caeultrasonics.com/fu-pagel.php3
99
Additional References
Harris D. A., "Light Spectroscopy", Bios Scientific PublisherLimited, Oxford, UK (1996)
Schroeder D. J., " Scanning Spectrometer of the Gillieson Type", Appl. Opt. 5, 545 (1966)
Schroeder D. J., "Optimization of Converging-Beam Grating Monochromator," J. Opts. Soc. Am.
60,1022 (1970)
Monk G.S., "A mounting for the plane grating", J. Opt. Soc. Am. 17, 358 (1928)
Gillieson A. H. C. P., "A New Spectrographic Diffraction Grating Mounting", J.Sci. Instr. 26,
335 (1949)
Namioka T., J. Opt. Soc. Am. 49, 951 (1959)
100
Appendix A
101
Al Machine Pictures
Light
Source
Temperature
controlled
storage station
Ultrasonic
cleaner
Circuit
board
Figure Al: The designed automated purification system for biologically active macromolecules
102
a) The light source
b) The cutting device and its transportation system
d) The circuit board
c) The cutting tip storage and cleaning station
Figure A2: Pictures of selected components
(See Appendix G3 for pictures of the temperature-controlled storage station)
103
A2 Parts List
Light Source
Part Description
Fig. #
Part Number
Manufacturer
Quantity
Deuterium Lamp (L2D2)
Power supply
Monochromator (100mm)
L6312-50
C4545
9030
Hamamatsu
Hamamatsu
Sciencetech Inc
1
Concave holographic grating
GR32/U
Sciencetech Inc
1
1
1
7292
Oriel Instruments
1
K45-340
Edmund Industrial Optics
1
K33-497
K54-011
4SQ-120BA34S
MC3479P
T22A-2518
00-166020
Edmund Industrial Optics
Edmund Industrial Optics
Thomson Airpax
Motorola
PIC Design
Schneider Optics
1
1
1
1
1
1
Q130125
A2047-ND
Esco Products Inc
Digi-Key
1
1
A2043-ND
WM4007-ND
WM2007-ND
Digi-Key
Digi-Key
Digi-Key
1
2
2
WM1205-ND
Digi-Key
1
WM1204-ND
Digi-Key
1
WM1000-ND
Digi-Key
3
WM1001-ND
Digi-Key
3
SW301-ND
Digi-Key
1
W410-X-ND
Digi-Key
100"
(250nm blazed 12001/mm)
Collimating Reflector
(57*38mm)
Mirror, UV coated
(75*75mm)
Mirror Mount (1.5inch)
45degree Adapter
Stepper Motor (1.8 degrees)
Driver chip
Zero backlash flexible coupling
Electronic Shutter
Prontor Magnetic 016
Window Fused silica (3*3",1/8")
D-sub Connector (Female, 9
Pins)
D-sub Connector (Male, 9 Pins)
.100" Center Header (9 Pins)
.100" Center Crimp Terminal
Housing (9 Pins)
Power Connector plug
(.062" Terminals, Panel mount)
Power Connector Receptacle
(.062" Terminals, Panel mount)
Power Connector Terminals
(.062" Terminals, Male)
Power Connector
Terminals(.062" Terminals,
Female)
Panel Mount Rocker
Switches(on/off)
Multi-Conductor Cable
(Non Plenum, 24AWG, 10
wires)
104
Light source
Part Description
Fig. #
Fan finger guard assembly
Part Number
Manufacturer
Quantity
CR287-ND
Digi-Key
1
BA1
PHl-ST
TRI
TR1.5
MB 12
61-3471
53-3505
Thorlab
Thorlab
Thorlab
Thorlab
Thorlab
Coherent
Coherent
2
3
2
1
1
1
1
1
1
2
1
1
(45 PPI, 40 mm square)
Base, fork, 1*3 inches
Post holder, 1 inch
Post, 1 inch
Post, 1.5 inches
Breadboard (12*12 inches)
Rail (19mm, 3")
Carrier (wide base)
Top cover
Front wall
Side walls
Back wall
Base
Bracket
Shutter holder
Heat sink
Lamp holder plate
Grating base
Grating holder
Grating shaft
LS101
LS 102
LS103
LS104
LS105
LS106
LS 107
LS108
LS 109
LS110
LS111
LS112
LS113
LS114
LS 115
LS116
Grating spacer
Grating arm
LS 117
LS118
1
1
Grating base legs
LS 119
LS120
1 each
Grating motor holder
Grating motor plate
Grating motor flexible coupling
LS121
LS 122
LS 123
2
1
1
Junction block
Inside wall
8
1
2
1
1
1
1
1
1
105
Cutting Device
Part Description
Fig. #
Part Number
Manufacturer
Quantity
Single edge stainless steel blade
Linear stepper motor
71980
L92121-P2
Electron Microscopy Sci.
Thomson Airpax
1
Driver chip
Linear slide
Linear slide
Rare earth magnets
Subminiature switches
Main frame
Small frame
MC3479P
BSP 10-45 SL
BSP 10-25 SL
64-1895
DH1C-BlPA
Motorola
IKO
IKO
RadioShack
Cherry Electric
ET101
ET102
1
1
1
5
3
1
1
Cutter holder
Cutter tip base
ET103
ET104
Stopper block
ET105
Flexible shaft plate
ET106
1
1
__5
2
________....
1
Cutter Transportation System
Part Description
Fig. #
Part Number
Manufacturer
Quantity
Stepper motor (1.8 degrees)
4SQ-120BA34S
Thomson Airpax
Driver chip
Power supply (12V, 3Amps)
48 Pitch rack (20 Degrees)
Spur gears, 1.25" pitch diameter
Anti-backlash pinion (48 pitch,
60 teeth)
AccuGlide miniature rail
AccuGlide miniature carriage
SS button head cap screws 6-32
MC3479P
2200503
AG-1
G5-60
AP48C-60
Motorola
RadioShack
PIC Design
PIC Design
Berg
1
1
1
4
1
1
RD-15-P-L760
CD-15-AA-A-P
92949A148-18-8
Thompson Industries, Inc
Thompson Industries, Inc
McMaster-Carr
1
1
25
Stainless steel cap screws 8-32
92196A193-18-8
McMaster-Carr
25
thread, 7/16" length
Reed contact switch
4900497
RadioShack
SX1515L-70
PIC Design
2
1
SCB3GL-15
SCB3GR-15
SH9-06
SCHI
SCA2-1515
SCH5
PIC Design
PIC Design
PIC Design
PIC Design
PIC Design
PIC Design
1
1
15
15
6
6
thread, %/"length
Aluminum extrusion
(1.5*1.5*70")
Corner brace left
Corner brace right
Button head screws (5/16,0.63")
Connector T-nuts (5/16")
Plastic end plate (1.5*1.5 Inches)
Pushlock fastener
Main Frame
Carriage frame #1
Carriage frame #2
TS101
TS102
TS 103
1
Rack
TS104
4
1
1
106
Cutting Tip Holder and Cleaner
Part Description
Fig. #
Part Number
Manufacturer
Quantity
4SQ-120BA34S
70155K57
Electrowave Corp
McMaster-Carr
1
21811-896
9008K21
9618K14
90291A108-18-8
VWR scientific
McMaster-Carr
McMaster-Carr
McMaster-Carr
1
1
1
10
CG1-2-A
Al-120
Berg
Pic Design
2
1
Ultrasonics cleaner
Box-Frame solenoids
(pull style, continuous duty,
12VDC)
Ultrasonic cleaning solution
Square aluminum Bar, 3/8"* 1'
Compression spring assortment
SS set-screws with nylon tip 440 thread, 3/8" length
Clamp- split type 1/8" bore
Precision ground shaft 1/8"
diam., 12" long, stainless steel
303
Holder base plate
Delrin arm
Shaft holder 1
Shaft holder 2
TH101
TH102
TH103
TH104
1
1
1
1
Bracket
TH105
1
Shaft
TH106
2
Temperature Controlled Sample Storage System
Part Description
Fig. # Part Number
Manufacturer
Quantity
Thermoelectric module
Module controller
6300/127/085A
5C7-350A
Ferrotec America
Oven Industries
2
1
Temperature sensor
Cooling fans + Heat sinks
Oven Industries
RadioShack
1
2
Silicone grease
Stainless steel pan
3/8-16,2" hex head cap screw
Polyurethane expanding foam
TS67-178
2730246;
2730248
2761372
4191T13
913009A632
8551KI1
RadioShack
McMaster-Carr
McMaster-Carr
McMaster-Carr
2
1
4
1
Swivel leveling mounts
6111K83
McMaster-Carr
4
107
A3 Vendors Information
Active Electronics,
73 first Street, Cambridge, MA
Tel:(617) 864-3588
Berg
499 Ocean Avenue, East Rockaway, NY 11518
Tel: (800) 232-BERG
http://www.wmberg.com
Digi-Key
Tel: (800) DIGI-KEY
http://www.digikey.com
Edmund Industrial Optics,
Tel: (800) 363-1992, Fax: (856) 573-6295
http://www.edmundoptics.com
Electron Microscopy Sciences,
321 Morris road Ft, Washington, PA 19034,
Tel:(215) 646-1566, Fax:(215) 646-8931
http://www.emsdiasum.com/ems/preparation/blades.html
Electrowave Ultrasonic Corporation,
Tel:(715) 426-7378 Fax: (715) 426-7351
Email: info@inter-netco.com
http://www.inter-netco.com
Esco Products Inc.,
171 Oak Ridge Road, Oak Ridge, NJ 07438
Tel: (800) 922-3726 , Fax (973) 697-3011
http://www.escoproducts.com
FerroTec America
40 Simon St. Nashua, NH 03060
Tel: (603) 598-7336, Fax: (603) 598-7272
Future Bearings,
15 Walkers Brook Dr, Reading, MA
Tel: (781) 942-9880,
http://www.ikont.co.jp/eg/product/product.htm
108
Hamamatsu,
360 Foothill road, P.O. Box 6910;
Bridgewater, NJ 08807-0910,
Tel:(908) 231-0960, Fax:(908) 231-1218
http://www.hamamatsu.com/
McMaster-Carr,
473 Ridge Road, Dayton, NJ 08810
Tel:(732) 329-3200, Fax:(732) 329-3772
http://www.mcmaster.com
Newark Electronics,
59 Composite Way, Lowell, MA 01851-5144,
Tel: 978-551-4300, Fax: 978-551-4329
http://www.newark.com
Oriel Instruments,
150 Long Beach Boulevard, Stratford, CT, USA, 06615-0872;
Tel: (203) 377-8282, Fax: (203) 378-2457,
http://www.oriel.com
PIC Design,
Tel: (800) 243-6125, Fax: (203) 758-8271
http://www.pic-design.com
RadioShack,
493 Massachusetts Ave, Cambridge, MA, 02139
Tel: (617) 547-7332
Schneider Optics,
285 Oser Ave, Hauppauge, NY 11788
Tel: (631) 761-5000, Fax: (631) 761-5090
Sciencetech Inc,
45 Meg drive, London, Ontario, Canada, N6E 2V2;
Tel: (519) 668-0131, Fax: (519) 668-0132,
http://www.sciencetech-inc.com/
Thomson Airpax,
Seven McKee Pl., Cheshire, CT 06410
Tel:(203) 271-6444, Fax:(203) 271-6400
VWR scientific,
Tel:(800) 932-5000
http://www.vwrsp.com
109
A4 Circuit Schematic of Selected Components
+12V
2
1
1K
3
MJ-30-55
GND
+12V
MC3479P
Stepper motor driver
+12V
L2
1
2
Li
13
GNDGND-
4
5
6
7
8
47K
GND-
CLK
2
1K
I
1
+12V
15L3
14 L4
13 -GND
12 -GND
11~
1n
CW/CCW
9
GND
MJE3055
3GND
4SQ-120BA34S
+12V
Q3
2
1K
2
3
I
14
MJE3055
3GND
I
RJC097
GND
+12V
-
I
0
a
CLK
Q
STEPPER
MOTOR
i
CW/CCW
2
1K I
Signal from computer
MJE3055
3GND
Figure A3: Schematic for the 4SQ-120BA34S stepper motor
110
Q
MC3479P
Stepper motor driver
+12V-
1
16 -+12V
L2
2
Li
3
4
5
15
14
GNDGNDGND
LA
Q3
11-
7
CLK
0 CW/CCW
9 GND
-8
2
3
I
RJC097
GND
CLK
CW/CCW
Signal from computer
Figure A4: Schematic for the L92121-P2 stepper motor
+12V
Signal from
computer
1
IK
iSolenoid
2
MJE3055
GND
Figure A5: Schematic for the solenoid
Signal to
computer
GND
4700
+12V
C
0
Q
STEPPER
MOTOR
L92121-P2
13 -GND
12 -GND
6
47K
Q2
L3
GND
+5Vy-
UA7800
Voltage regulator
Limit switch
sensor
Figure A6: Schematic for one of the feedback sensor
111
Q4
Appendix B
112
B1 CCD Camera Specifications
D'
A -
A
$
I
H
T
EFE
The SpectruMM:250B is a high-performance digital camera syslen featuring a Hamamatsu back-illuminated spectroscopic-fomnat CCD. The
1024 x 250 imaging array isideal for general-purpose spectroscopy Back-illumination and thermoelectric cooling to -35 C give the
SpectruMM:250B the sensitivity and low noise necessary for Raman or weak-fluorescence applications. Its 6-mm height and full 24-mm
sprectral coverage deliver multislripe capability as well.
E
A T
U R E
S
N E
FIT
S
Hamamatsu CCD sensor
Delivers industry-standard performance
1024 x 250 imaging array
24 x 24-pm pixels
6-nm-tall imaging area
Ideal format for general-purpose spectroscopy
Provides good resolution and excellent full well capacity
Wdeal for rapid, multistripe spectroscopy
Back-illuminated CCD
Offers higher sensitivity for low-photonflux applications
Optional dual digitizers
High speed provides rapid spectral acquisition
[ow noise provides the bes signal-to-noise ratio
Easy yet sophisticated Windows GUI controls and integrates
camera with spectrometer
Automates data acquisition, analysis, and display
WinSpec
SpectruEMM
113
I
"ID,
A
-T
100
90
H
f
EI
A
1 -T
05N
-I-A-
80
t"
S
A,
70
-
-
- -
-
- -
-______
60
50
40
30
C.13
20
10
0
20 0
600
400
1000
800
1200
Wavelength (nm)
IS
P
EF C
CCD image sensor
CCD fomiat
I
F
I
C
Hamamatsu; scientific grade; MPP; back- illuminated:
available with Uenhancernent coating
1024 x 250 Imaging pixels; 24 x 24pm pixels
100% fill factor; 24.6 x 6 0mm imaging area
Speclrometr well capacity
550,000 e
CCD read noise
Be
System read noise
10 e rms @ 100 kHz;
30 e rms @1 Wz
Nonlineamy
Nonunifornity
Dynamic range
<2% for 16 bits
<±3%over entire CCD area
16 bits @ 100 kHz;
12 bits @ 1 MHz
Scan rate
100 kHz o 1 MHz
Vertical shift time
Spectral rate
Dark current
Operating temperature
Thermostating precision
nrs @150 kHz
10
50 Hz, full-vertical binning, 100kHz digitization;
200 Hz, full vertical binning, 1-MHz digitization
<15 e-/ps @
-30 C
-35 C wth forced air circulation
±0.04 C over entire temperature range;
dark-charge stabilized to ±06%
EjrMM
JbticmmJ
RSbtmin 4wi
ROPER SCIENTIFIC
i
PRINCETON INSTRUMENTS
114
o
(
mi)
1
(trnf-
I1'
-NOTE: T-IIKON MOUNT ADAPTER AVAILA8L
Lens Specifkafion
Lens construction 3group 3eiement (al QVARTZ LENS)
256m0
Wave length
77.55mm* 5X
Focal lenglh
7131mmt 5K liA air)
Back focal
17.526mo C-mowt
Frnge back
F/3-8 *5% - F/22
Aperture ratio
18.0
:
hoage circle
Filter Thread
Angle of view
knage to object distance = infinfy
Diagonal
- 6.64'
OPT Distortion: -0.27% MLiagonal)
: -0.3x
Magnification
Image to oIJect distance = 436.8 mm
Working distance: 204 mm
OPT. Distortion: -0,13% (agonad)
023.0
Aperture of front lens
210
Aperture of rear lens :
Coating
Al surface are
antiretledion coated.
tMgFZ single coat)
Mounting
:
-mount & T-mount0 49 me P=0. 75
FM ter thread:
049 P=.75
1
T-
ONT Thread
K142.0 P=0.75
(ff117 at INF
';26
at INFIll.4}
9
*56.O
053
I
U
I
U
A
Z*
P4
I
L
(101.5) at Hag = -0.3x
138.91 at
54.9
Ii i.
=-3x
4ag
UNJVERSE KOGAIU (AMERCA), INC.
111 MSVKf.
OViMEAa
VOWK t&77
TiTLE
78mm F/3.8 UV IIMAGING LENS
DATE
912/99
SCALE
None
NO.
UV8O40B
0RAWN
AO
B2 XY Stage Specifications
Figure B: 402 LN Series XY Table from Daedal. [4]
Spe cifications
4020CO
Travel
Life @rated specifiation X1 rnilln inches (mm)
Positional Accuracy xO.CX)1 in (mm) Std Grd
Prwc. Grd.
over table travel
6
10
2.9
0.46
(15Q
f250
(75)
f12)
Positional Repeatability x o~om im I mm) Sid Grd.
±0.46
(±12)
Prec. Grd_
Straight Line Accuracy x0.001 in (mm) Std Grd
Prmc. Grd
Ovr Total Table Travel
Flatness Accuracy E0.C01 in (mmli Std Grd
Over Total Table Travel
Prec. Grd.
Std Grd
Max Screw Speed Orps
Prec. Grd
Max Acceleration Din/5ac jinwtwe. Std Grd
Prec. Grd
Duty CyclL % a raiuto Lmei pe eyLE Std Grd
Prec. Grd
Norrnol
Drect Loading" DIbs (kg)
±0.07B
0.93
0.31
0.93
Inverted
Lc d per BerOng
DIb5 (kgt) Both grades, Nbrmnal
Axial Loading Dibs (kg)
Inverted
Std Grd
Prec. Grd
Input Inertia" D10 z-in-sec 2
Maimum Running Torque Dcz-in (N-m
Maximum Bneakawa.y Torque Doz-in fN-ml
Drive Screw EfficiencyEtd &Prec Leadscrew D%
Coeffrziant of Lirear Bearing Friction
Carriage Weight Dlbs (kgfl
Longitudinal Span between Bearing Inuckenter
Lateral Span between Bearing Rail Centers
Bearin RAil CEn ter tp Caniao Moun ing 5ur fa
Tnha Winht - Ih- lkin
116
(2)
(24)
(B
(241
(B)
0.31
15
25
386
3B6
(9BC)
(9KDC
50
75
160
40
40
10
10
2B
0.912
12
13
(72,7)
f18,2)
(1B,2)
(4,.5)
(4,5)
(12,71
Q.5B951
(0,(3471
U.093 21
30
0.01
0.14
1 .48.5
1.456
(37,Q
0.541
(13.71
1 12
11 471
(.0
(37,7)
Miniature Linear Positioning Systems
.- .
.
402000LN Series Dimensions
1awpIikr
USX 11s
4599
ao
40f
" gn
c'
I
T
see odicna
k
M51IM
Ilf ddalf,
57.7
n
155
n
ri
at a
I
1.
NEMA
23
mCit
toadlw MSX a
U
4nfr~lhib.
4 ha"s
NEMA t79
-A
mowt
555995
-~-
1
fho
Mf1,
6r4
Mrf.
4
Im
117
WIwit
-4-
arsi d NE VA 23 r11 f9f1i5A
--
Appendix C
The Light Source
C1 Assembly Drawings
118
8
II
7
6
THE INFORMATION CONTAINED IN THIS DRAWING I THE SOLE PROPERTY OF
MIT and Alpine Phamaceutical ANY REPRODUClON N PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIITED.
5
1
8 7
16 4
3
I
2
I
I
D
D
2
'I
-~1
0
I
4
c
c
1
-
CD
4-
attached to blocks by
3
4x 6-32,1/4" button head screws
Front wall
attached to blocks by
4x 6-32,1/4" button head screws
Cover
4
4x 6-32,1/4" button head screws
Back wall
2
B
Side walls
AI
-
B
attached to blocks by
attached to blocks by
4x 6-32,1/4" button head screws
5 Wall block top (3 sides tapped)
5* Wall block bottom
(clearance hole vertical, attached to base
6
A
ITEM
NO.
by 4x 6.32,11/16" pan head phillips screws)
PART
NO
IDER
OR
IENEYO
O.OR
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE 14 INCHES
TOLERANCES ARE:
FRACTIONS DECMALS ANGLES
+
.XX+ .O1 + 1
S.xxx.iooS
Heat sink face down
(attached to cover by 2x 2-56, 3/8" cap screws
with a #2 washer between cover and heat sink)
7 Window (attached to cover with silicon sealant)
-Mr.LUtSPFAIRL
DESCRIPION
SNO.
IS
PARTS LIST
CAD GENERATS) DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
1I
/
/1
I
66
1I
A
Eric Hoarau March 2001
Light Source
Assembly outside walls
RESP RUG
b11
SAE
USED ON
DO NOT SCALE DRAWING
APPLICATION
ts
MIT Mechatronics Research Laboratory
CHECKED
FINISH
NEXT ASSY
9
If
4
4
I
1
REDD
DATE
MATERIAL
Aluminum 5052
PECIFCAION
PUAL
i
j
SCALE
DWG.
LS001
NO.
2 I CAD
y
FILE:
SI3
U
OFl
8
II
6
5
43
II
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OP
MIT and Alpine Phamaceutical ANY REPRODUCON IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHMITED.
2-I
-I
D
D
00
000
000
000
-o-
8
c
c
o®o
0
0
~:
US
0
0
*0
0
A
1 Grating assembly
(bolted to base by 2x 1/4"-20,1/2" scre ws
+ 1/4"washers)
2 Lamp assembly
(bolted to base by Ix 1/4"-20,1/4" scre w)
3 Fan + Filter
(bolted to base by 2x 6-62,7/8" screws )
4 Inside wall
5 Collimating assembly
(bolted to base by Ix 1/4"-20,5/8"scre w
+ 1/4"washer)
6 Mirror assembly
(bolted to base by Ix 1/4"-20,5/8"scre
+ 1/4"washer)
7 Brackets for inside wall
(bolted by 2x 6-32,3/8"screws+ #6 washers)
8 Electronic shutter
9 Terminal block
(bolted to base by Ix 6-32,3/8" screw)
10 hutter holder
(bolted to inside wall by 2x 6-32,3/8" sc rews)
11 Slit
(holded in place by I x 4-40,3/16"screv + #4 washer)
B
3
A
ITEM
NO.
PART OR
IDENTIFYINGNO.
NOMEN.LAIU~t
MATRIAL
SPECICATION
OR DESCRIPTION
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS DECIMALS ANGLES
+
.XX+ .01
+
.XXX+.005
1
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
MIT Mechatronics Research Laboratory
DATE
APPROVALS
A
Eric Hoarau March 200
Light Source
Assembly of components
CHECKDW
MATERIAL
Aluminum 5052
Q Y
REQD
UST
I
RESPENG
FINISH
NEXT ASSY
USEDON
APPUCATION
I DWG. NO
.
CAD
SCALE ICAD
SCALE
SE
AE
I DO NOT SCALE DRAWING
:3
L
LS002
PILE:
FILE:
IISHEET
SHEET
I
RRV.
OP
OF
I I
8
6
7
THE INFORMATION CONTAINED R4 THIS DRAWING IS THE SOLE PROPERTY OF
MIT and A wN Phamaceutical ANY REPRODUClON I PART OR WHOLE WITHOUT
WRITTEN PERMISSION S PROHMITED.
5I
4
III
3-I
2
-I
12
6
D
D
--9
5
9
FE
CIO
o-
3
10
c
FE
7
0
-I
7
0
0
9
FE
FE
FE
0
A
45*Mirror Assembly
0
0
FE
0
A
1 Rail (2.5")
2 Rail Block (1")
3 Postholder (1" length)
(bolted to base with a 1/4"-20,3/8" bolt)
4 Post (1" length)
5 Lamp base
(bolted to post with a 8-32,3/8" bolt + #8 washer)
6 Deuterium lamp
(bolted to lamp base with 2x 4-40,3/8" bolt +#4 washe
7 Base fork
8 Post (1/2" length)
9 Mirror + Minimount + 45'adaptator
(attached to post with a 8-32,5/8" set screw)
10 Post holder (1.5" length)
(bolted to base with a 1/4"-20,3/8" bolt)
11 Post (1" len th)
12 Collimator model 7292)
(attached o post with a 8-32,5/8" set screw)
B-
B
Collimator Assembly
Lamp Assembly
ITEM
S
PART ORSPMItKA
IDENTIY
N
SPECIHCAPON
N
I
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFED
DIMENSIONS ARE N INCHES
TOLERANCES ARE:
FRACTIONS DECIMALS ANGLES
+
.XX+ .01
+ I
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
CHEOCKED
Aluminum 5052
RESPENG
Light Source
Assembly Components #2
MATERIAL
FINISH
NEXT ASSY
I
SIZE DWG. NO.
USED ON
APPLCATION
A
March 200
NHoarau
.xxx+.00S
MIT Mechotronics Research Laboratory
DATE
DO NOT SCALE DRAWING
UAL
t:NU
3
SCALE
SCL
LS003
FILE:
ICAD
CADPPLE:CTEETOO
IRE-V.
SHEET
I
OF
I1
8A
I
7
7
66
I
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY
MIT and A pine Pharmaceutical ANY REPRODUCION IN PART OR WHOLE
WRITTEN PERM5SION IS PROHNITED.
I
OF
4
I
5
3
I
2
I
1
WITHOUT
D
D
13
1
4
13
r)
c
US
0
-
c
1
7
E
0
-
3
1 Base
B
2 Miniature switch
3 Grating arm
S'
RE
(bolted to grating shaft by
and a plastic washer)
1x 8-32,3/8" cap screw
CD
4 Counter
5 Base leg #1
(bolted to base by Ix 1/4-20, 1/4" cap screw)
6 Base leg #2
(bolted to base by Ix 1/4-20,1/4" cap scre w)
A
B
2*
6
(bolted to base by 2x 0-80,3/8" cap screws)
PART
ITWI
SPECIFIED
LE
TOLERANCES ARE:
FRACTIONS DECIMALS ANGLES
+ 1
-Xx+ .01
+
.XXX+.005
MATERIAL
Aluminum 5052
A
N
DAT E
NHoarau MarCh
C
CHECKED
RESP NG
MIT Mechatronics Research Laboratory
NUALLY UPDATE
APPROVALS
R
SPCC
OR DESCRIPT
PARTS LIST
NOHERWISE
U
ARE IN INCHES
DIMVENSIONS
10 Flexible coupling
11 Grating holder
12 Grating shaft
13 Bearing
OR
IDENIFYIG NO.
No.
7 Motor holder bracket
(bolted to base by 2x 4-40,3/8' cap screw)
8 Motor holder
(bolted to motor bracket by 2x 4-40,3/8' c ap screw)
9 Motor
(attached motor holder by 2x M3 nut)
5
200'
A
Light Source
Diffraction Grating Assembly
FINISH
14 Spacer
15 Concave grating
NEXT ASSY
USED ON
APPLICATION
I
IES
I
~ ~ NOGCAEDRWN
AL N
DO NOT SCALE DRAWING
DO~~
4,.
A
I
'
|
Krv
A]'"
" LS004
I
ISCALE
SCALE
CAU
,C(AU
2
TILL:
MtE:
I
SHEET
OF
'
C2 Parts Drawings
123
8
7
6
4
5
3
1
422
i5
THE INFORMATION CONTAINED IN THIS DRAWING T5 THE SOLE PROPERTY OF
MIT ond Adpine Pharmaceutical ANY REPRODUCION W PART OR WHOLE WITHOUT
WRITTEN PERMISSION 5I PROHIBITED.
D
D
9.500
11.300
(10.550)
6
.375
0
0
4 x 0.150 THRU
LJ 0 .275T.075
-
1.272
c
1.272
c
-
12. 00
02.50 THRU
( i.250)
2.50
LJ 2-56 TAP
5. 00
US
0
B
EIIZ
LJ 03.25 T0.120
3.625
0
W.125
0~
B
0
.750
Bottom View
Top View
ITEM
NO.
PART OR
IDENTIFYNG NO.
NMN
OR DESC
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE N INCHES
TOLERANCES ARE:
FRACTONS DECIMALS ANGLES
+
.XX+ 0O1 + 1
A
Plate has a 3/16" thickness
xxx+.oO5
CADGENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
SPEC A
N
MIT MeChatroniCs Research Laboratory
DATE
A
6RAW
E"c Hoarau March 2001
Light Source
Cover
CHECKED
Aluminum 5052
Z
RED
MAT
LIST
RESPENG
FINISH
NEXT ASSY
USED
ON
oI
I //
I|6
Es
5
SCAE
DO NOT SCALE DRAWING
APPUCATION
T
1~
I
I
QUAL ENG
.5
j
SCALE
VAD-
|CAD
LS 101
FILE:
SHEET
I
OF
8
i
5
4
2
3
THE INFORMATION CONTAINED IN THIS DRAWING is THE SOLE PROPERTY OF
MIT and Alpine Pharmaceutical ANY REPRODUCTON N PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
11.750
.250
L
-(11.250
~1
.250
c
c
4x 0.150 THRU
(4.225)
C
4.725
r
pil
-o.880
-0
B
4-
.996
01.024 THRU
1
ITEM
NO.
PART OR
IDENTIFYNG
B
MATERIAL
NU"INLAU
SPECIFICATION
OR DESCRIPTON
NO.
QTY
REOD
PARTS LIST
Plate Thickness: 1/8"
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE N INCHES
TOLERANCES ARE:
FRACTIONS DECIMALS ANGLES
+
.XX+ .01
+
A
1
.xxx+.00S
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
MIT Mechatronics Research Laboratory
DATE
APPROVALS
A
Feb 2001
EriC Hoarau
Light Source
Front Wall
CHECKEO
Aluminum 5052
i
RESP ENO
FINISH
NEXT ASSY
USED ON
SIZE IU.
f
Y
I
WU.
QUALENG
DO NOT SCALE DRAWING
APPLICATION
I'3 0
I
SCALE
ICAD FILE:
LS 102
ISHEEI
8
8
53
A.
4
1
3
2
IN
THE INFORMATION CONTAINED
THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Apine Pharmaceutical ANY REPRODUCTON IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHBITED.
D
D
11.300
.375
.250
c
F
c
4x 0.150THRU
4.
-
I
10.550
4.224)
-
4.724
B-
1
nr
B
0
C=
.125
I PART OR
ITEM
NO.
IDENTIFYR
MA ERIAL
SPECIFICATION
NuMtNU
OR DESCRIPTION
GNO.
4
REQD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS DECMALS ANGLES
+
.TX+ .01
+1
.555+.005
Plate Thickness : 1/8"
2 Parts are needed
A
CAD GENERATED DRAWING
DO NOT MANUALLY UPDATE
RESP ENG
Light Source
Side Wall
i
FINISH
NEXT A SSY
8
|6
I
SAE
USED ON
APPLCATION
A
Eric Hoarau March 2001
CHECKED
MATRIAL
Aluminum 5052
MIT MechatronicS Research Laboratory
DATE
APPROVALS
DRAWN
DO NOT SCALE DRAWING
4
I
MAL EC
DWG. NO.
SCALE
T
ICAD FILE:
LS1O3
15Httl
OF
I
8
1
7
6
1
6
8
5
4
L
1
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
RT End
APE P
PRoceuticd
ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERNMSON IS PROHIBITED.
33
2
1
1
11.750
11.250
.625
D
(.969)
250
r- .250
4x
.508
4
D
.150THRU
4.225)
.756
00
-.
(1
DETAIL A
SCALE 1 : 1.5
c
4.725
o[--o 0
1.500
~0
0-
.800
.650THRU
L.J .800Wf.065
.650
L
c
0
.265
B
0.125
--
-
B
Plate Thickness: 1/8"
.984 -..
750
0.375
1.750
.425-
.963
1.000
.750
A
ITEM
N
0
-
FART
NO.
SPECIFICATION
UNLESS OTHERWISE SPECIFIED
IMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
-
+
DECIMALS
XX+
.XXT4+=5
CAD GENERATED DRAWINGE
DO NOT MANUALLY UPDATE
APPROVALS
ANGLES
.. +1
MIT MeChatroniCs ResearCh Laboratory
DATE
A
ErC Hoarau Feb 2001
Light Source
Back Wall
CHECKED
MATERIAL
Aluminum 5052
DETAIL B
SCALE 1 : 1
6
1
1
6
FINISH
MW
NET ASSY
I
I
RED
PARTS LIST
L
.50- - 2.000
NMATERAL
ESCRIPTN
OR
RTYING
APPLICATION
SAE IWG
NG
ugA-
USEDON
NG
SCALE
IDO NOT SCALE DRAWING
4
3
NO
LS 104
ISHEET
CAD FILE:
I
I
OF
5
1
6
8
4
IL
3
1
I
2
1
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Apine Phcrmaceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
11.300
(.375)
D
( .375 )-
--
10.550 )
'0000
-
T-
12. )00
(
D
.5
oo
0000
4x 6-32 TAP THRU
( .375 )
5x
C
75
(-750)
16 x 0.200 THRU
.300 SPACED
4-20 TAP THRU
1.125
!50)
C
0
8.125
8.250
0
7.750
00
0'
0
5.898
00
0
-FT
-9
B
0
B
1.500)
i
3.5006.063
8.917
10.000
ITE
PART
. AT
ORNOMENCLATURE
DESCRIPTION
IOR
NO.
M.ATER AL
SPECIFPCATON
WIT
REDo
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DMENSIONS ARE IN INCHES
TOLERANCES ARE:
Plate thickness: 3/16"
FRACTIONS
A
DECIMALS
.XX+ .O
.xxx+m5s
+
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
ANGLES
+1
MIT MeChatronics ResearCh Laboratory
DATE
APPROVALS
A
ErIc Hoarou March 2001
Light Source
Base drawing #1
CHECKED
MATERIAL
Aluminum 5052
RESP ENG
FINISH
NEXT ASSY
ON
5
I
/
I
6
I
5
SIZE
DO
APPCATION
9
QUAL
NOT SCALE DRAWING
4
i
DWG NO.
A
ENG
-r
SCALE
-
LS105
CDFL:SET
2
I
O
8
7
1
6643
1
~i
7
I
INFORMATION 8
CONTAINED
IN THIS DRAWING IS THE SOLE PROPERTY OF
I i THE
MIT and Apine PhamaCeutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
4
55
3
WRITTEN PERMISSION IS PROHIBITED.
1
2
7.031
5.020
0
D
0000
0000
0000
2.646
0.080T.08
D
0000
-,1
1.375
0.080
3.937
CT
.080
6-32 TAP THRU
7T
3.701
2.000
0.080T.080
C
C
61.540
6-32 TAP THRU
4.773
0.080T.080
\
6-32 TAP
Z
01.400
)
(6.969
0.080T.080
8.750
6.248
4-
4.000
0
2.500
0
0a
1.750
Cr
:0
B
1.457
2x 6-32 TAP THRU
(.6:4
---
2.220
B
2.275
-5.500
6.500
8.000
TEM
Plate thickness: 3/16"
A
X)PART 'OR
ENIFNG
S
AL;LIUI
SPECIICAN
OR DESCRIPTION
NO.
RED
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
A
FRACTIONS
DECIMALS
+
.XX
DI
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
ANGLES
+1I
MIT Mechatronics Research Laboratory
DATE
A
Eric Hoarau March 2001
Light Source
Base Drawing #2
CHECKED
MATERIAL
Aluminum 5052
ESP ENG
FINISH
NEXTASSY
USEDON
DO NOT SCALE DRAWING
APPLUCATION
8
7
1I
6
1
5
1 '.
4
I2UAL
SIZE
DWO.
SCALE
1 ,
NO.
ENG
13
|
LS106
SHEET
CAD FILE:
2
I
1
OF
8
7
5
1
6
4
L
1
3
1
2
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Ampine PharmaCeutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
8 SQUARE BLOCKS NEEDED
*
4 are 6-32 tapped on all sides
*
4 are 6-32 tapped on 2 sides
and 6-32 clearance hole on third side
6-32 TAP THRU ON 2 SIDES
6-32 CLEARANCE THRU ON THIRD SIDE
C
R.R 1
o
US
0
0
Q
c
6-32 TAP THRU
ON ALL 3 SIDES
.500
.500
B
L1
B
.500
0
.500
--
-
Top Wall Block
Bottom Wall Block
PART
ITEM
N
ATERAL
SPECIFICATION
NOECAL
OR DESCRIPTION
OR
IDENTIFYNG
NO.
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
+
A
DECIMALS
XX+ .T
.XXX+Ml5
ANGLES
+T
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
MIT Mechatronics Research Laboratory
DATE
A
Ecri Hoarau Feb 2001
Light Source
Wall Blocks
CHECKED
MATERIAL
Aluminum 5052
RSP ENG
FINISH
NEXT ASSY
ZE DWG. NO.
USED ON
LS107
AE
a
7
I
6
1
APPLICATION
5
rUAL ENG
DO NOT SCALE DRAWING
4
1
SCALE
3
I
CAD
2
PILE:
1
SHEET
1
S REV.
OF
8
I
I
I
.
7
1
1
6
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine PharmceutiC ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
I5
1
4
5
CF
D
6-32 TAP THRU
1
.L
3
I
1I
22
D
0
4.724
-2.375
=
.625
C
c
.9
S
2.028
0
(2.417)
B
5.465 -------
--
B
.625
3.878
6-32 TAP THRU-
2.402PART OR
IDENTIFYING NO.
ITEM
1.315
NO.
-'1.25
-
MATERIAL
SPECIFICATION
NOMENCLATURE
OR DESCRIPTION
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
A
FRACTIONS
DECIMALS
+
.XX
Dl
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
ANGLES
+1I
Plate Thickness: 1/8"
ErIC Hoorau
MIT Mechatronics Research Laboratory
DATE
Feb 2001
Aluminum 5052
NEXT A
SSY
FINISH
USED ON
i APPLICATION
I
/
I
6
5
.
RESP ENG
MAL
NU
SIZE
DWG.
A
rAL ENG
I DO NOT SCALE DRAWING
4
I
NO.
CAD FILE:
SCALE
I
A
Light Source
Inside Wall
Drawing #1
CHECKED
MATERIAL
6
REOD
PARTS LIST
I
LS 108
SHEET
I1
I"Ev
OF
8
I
7
6
I5
I THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine Phrs cuticmi ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITEN PERMISSION IS PROHIBITED.
1
5
4
A
11
22
2
3
I
1
1.160)".
I
.872
D
D
(1.77)1- -o
0
0
-4-40 t pped
.925
2.756
0.591
Oil
C
.591
-(
-
.925
DETAIL A
SCALE 1 : 1.5
C
2x 8-32 Clearance hole
?25
0v
FE
B
1~
I
0
0
B
4.724
( 2.417 )
NO.
,. -0
NOINCLIU
PARTS
.126
A
'RACTIONS
+
DECIMALS
XX+ DI
.XXX+fl05
.079
ANGLES
+1I
NEXT ASSY
-.
1
0
RESP ENG
DO NOT SCALE DRAWING
IQUAL ENG
A
Light Source
Inside Wall
Drawing #2
Eric Hoarau Feb 2001
CHECXED
I
FINISH
SE IDWG. NO.
USED ON
APPLICATION
0
MIT MeChatroniCS Research Laboratory
DATE
APPROVALS
Aluminum 5052
RED
UST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
MATERIAL
DETAIL B
SCALE 1 : 1.5
SPECIFICATION
OR DESCRIPTION
TPINGRNO.
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
1A RA111
I
I
ICAD
SCALE
2
LS1O9
FILE:
ISHEET
OF
8
7
,
6
5I
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Arlne Phwrrnceutica ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
4
3
1
2
-
.156
D
ILIZZI
.750
.297
.200
c
-
.500
c
125
.200
H1
0-
0
.500
1.000
B
.125-
B
.156
1.000-
ITEM
NO.
-
FART OR0MENCLATURE
OENTIFTNG NO.
-
A
FRACTIONS
+
MATERIAL
DECIMALS
MAT
XX+ 0l
.XXX+.005
APPROVALS
+
1
SPECIFICATION
TY
REQD
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
ANGLES
- .300
OR DESCRIPTION
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
-
|
MIT Mechatronics Research Laboratory
DATE
A
Eric Hoarau March 2001
CHECKED
Light Source
Bracket for inside wall
Aluminum 5052
FINISH
NEXT ASSY
USED
ON
d
I
/
1
6
1
5
764
I CAD
SCALE
I
'
'
1
'
LS110
N
A
'DUAL ENG
DO NOT SCALE DRAWING
APPUCATION
2
FILE:
SHEET
1
OF
8
I
I
7
5
6
4
3
1
1
1
2
2
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine Pharmaceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
0
.250
n
-r
0
SECTION A-A
SCALE 1 : 1
c
1.850
2x 8-32 TA P THRU
0
CD
A
A
US
C
2 .165
C,
CD
US
B
1.850
0 1.300 THRU
CD
LJ 0 2.087T.177
0
CD
N
SPECIICATION
OR DESCRIPTON
TIYNGNO.
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
2.165
A
FRACTIONS
+
DECIMALS
ANGLES
.XX+ .T
+ 1
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
DRAWN
MIT Mechatronics Research Laboratory
DATE
A
Eric Hoarau Feb 2001
Light source
Shutter Holder
CHECKED
MATERIAL
Aluminum 5052
FINISH
NEXT ASSY
USED
APPLICATION
8
1
7
T
Mi-
ON
DO NOT
RESP ENO
SCALE
DRAWING
tNG
8UAL ENG
SIZE
DWG. NO.
:V
LS111
A
CAD
SCALE
2
SHEET
FILE:
|
1
OF
8
I
.
Ii
6
,
4
..
I5
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Apin, PharmOceulicol ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
3
I
I
2
I
D
D
- .236
1.772
.250
1.272
C
.250
2 x 0.110 THRU
0
1.772
1.272
pr
--0
PART ORNOMNCLATUR
IDENTIFYING NO.
o.
OR
MATE
SPECIFICATON
DESCRIPTION
LIST
REGD
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
+
A
DECIMALS
.XX+
D
ANGLES
+ 1
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
MIT MeChatroniCs ResearCh Laboratory
DATE
A
EriC Hoarau April2001
Light Source
Heat Sink
=NECKED
MATERIAL
Aluminum 5052
tESP ENG
FINISH
NEXT ASSY
APPLICATION
I1
66
I
SIZE
USED ON
DO NOT SCALE DRAWING
LA
SCALE
DWG. NO.
ICAD
IRtV.
LS1 12
FILE:
I
I
SHEET
I
I
Of
I I THE
6
7
8
INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine Ph=rmaceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
5
1
8
A.
6I
4
i
3
2
I
I.I
D
D
3.18
52 -----
0
0
8-32 Clearance ho le
C
C
7
P9.
I-
( 38)
3x 4-40 TAP THRU-
65
PD
B
B
23
26
e9
ITEM
NO.
23
OR
23
Al
FRACTIONS DECIMALS
+
XX+.1
ANGLES
+
1
MIT Mechatronics Research Laboratory
DATE
Eric Hoarau Sept 2000
CHECKED
MATERIAL
Aluminum 5052
FINISH
NEXT ASSY
SED ON
RESP ENG
MvFG
LNU
Size
00
/
I1
I
i
b
DO NOT SCALE DRAWING
Light Source
Base for
Deuterium Lamp
LS 113
DWG. NO.
A
APPUcAICON
y
REGO
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
SPECIFICATlON
DESCRIPTION
PARTS
UNLESS OTHERWISE SPECIFIED
DOI1NSIONS ARE IN ?Mileter
TOLERANCES ARE:
-*-~ 26
A
PART OR
DENTIFYING NO.
SCALE
ICAD
FILE
I
SHEET
I
A
[REV.
OF
. I THE
8
7
6
5
1
4
.L
i1
INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
nd Alpine Phomoceudlcal ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
|1
33
I
22
WIT
II
D
L- 2.250 -
9
D
.350
--
R.250
2.800
K1*
i
1.500
00
0
9
\0
POCKET CUT
L- W.625
4.900-
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0
1.750 -
-
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10
0
=
9
0
130
-
- .290
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0
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1.120
c
.3
-.
.900
f-
P..,
4x 4-40 TAP
0
9
0-
1.220
W.500-----B
0.
This part is from the 9030 monochomator
- Base is cut to the specified dimension
- Pocket is machined on the top
- Four tapped holes are machined on the side
- Two tapped holes are machined at the bottom
-I
9
eD
DETAIL A
SCALE 1 : 1
-
.850
j
9
P
ITM
O.
PS
MATERAL
R ORMN;LT
ITING NO.
-
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
1.000
FRACTIONS
DECIMALS
ANGLES
+
.XX+ '0l
+1I
SPECI
ESCRPTION
PARTS
S
1.000
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
.185
FINISH
NEXT ASSY
2x 2-56 TAP T.500
S
7
I
6
Aluminum 5052
MIT Mechatronics Research Laboratory
A
Eric Hoarau March 2001
APPLCATION
I
5
Light Source
Grating Base
RESPr
ENG
-
USED ON
DO NOT SCALE
4
DRAWING
D
DATE
APPROVALS
DRAWN
CHECKED
MATEIRIAL
T
ATTON
LIST
SIZE I DWNG.
NO.
SCALE
CAD
3
T
2
1"-
LS 114
rUAL ENG
SHEET
FILE:
I
'
OF
7
8
6
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine PhamoceutiAl ANY REPRODUCTION IN PART OR WHOLE WITHOUT
II
1
8
WRITTEN PERMISSION IS PROHIBITED.
~~ 5
16
4
1
3
1
2
I
I
D
D
1.000
11
E.
'i
IC
c
c
0c
2.000
This part is from the 9030 monochromator
The arm at the base is cut from the original
part to obtain the drawn part
-0
B
B
sED
0
PART ORNOME
OENTIfYING NO.
ITEM
NO.
1.250-
MA ERIA
OR
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS
FRACTIONS
ARE IN INCHES
A
+
MATERIAL
DECIMALS
.XX+ .OT
.XXX+.005
ANGLES
+1I
DRA
APPROVALS
WN
EID
RED
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
TOLERANCES ARE:
SPECIFICATION
DESCRIPTiON
MIT MeChatroniCs ResearCh Laboratory
DATE
A
EriC Hoarau March 2001
Light Source
CHECKED
Grating Holder
AlUminum 5052 RESP ENOG
FINISH
NEXT ASSY
APPLICATION
1
1
1
6
1
I
5
SIZE
USEDON
DO NOT SCALE DRAWING
11
DWG.
NO.
CAD
SCALE
z2
LS115
FILE:
SHEET
OF
8
I
a
Ii
5
i
i1
4
THE INFORMATION COTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Apine Ph=rnceutica ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
|1
33
22
1
D
D
This part is the grating shaft from
the 9030 monochromator.
The shaft lenght is reduced
from the threaded side of the shaft
The threaded hole is then retapped.
40
9e
c
c
Li
8-32 TAP ~.600
1.150
.025
4-
0a
n
-
B
B
0@
G.250
PART OR
IDENTIFYING NO.
ITEM
NO.
MATERIAL
SPECIFICATION
NOMENCLATURE
OR DESCRIPTION
Quf
RED
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTONS
A
+
MATERIAL
NEXT ASSY
I1
//
I
66
b
ANGLES
.XX+ .01
+1I
S4
A
Eric Hoarau March 2001
Light Source
Grating Shaft
Aluminum 5052
E.... --
DO NOT SCALE DRAWING
t
MIT Mechatronics Research Laboratory
DATE
APPROVALS
CHECKED
USED ON
APPLCATION
ts
DECIMALS
XX+WSI
.xxx+.oo5
CAD GENERATED DRAWING,
DO NOT MANUALL Y UPDATE
4
1
SIZE IDWG. NO.
SCALE
1I
j
j
ICAD
z
Rev.
I
LS1 16
A
FILE:
SHEET
I
I
I
I
OF
8
5
1
L
1
4
3
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Aipine PhrmceutiCd ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
.330
0
0
"J
c
0.625
0
This part is the grating shaft bearing spacer
from the 9030 monochromator.
The spacer lenght is reduced to .330"
B
0.400
rjs
WB
ITEM
NO.
PART OR
IDENTIFYING NO.
SPECIFICATION
OR DESCRIPTION
Ulf
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
(B
MATERIAL
NOMENCLATURL
A
FRACTIONS
DECIMALS
+
.XX+fl
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
ANGLES
+1
MIT Mechatronics Research Laboratory
DATE
A
RAWNr
EriC Hoarau March 2001
Light Source
Grating shaft
CHECKED
Aluminum 5052
;USP
-END
bearings spacer
FINISH
rNEX ASSY
APPUCATION
a
1
8 I
77
SIZE
USED ON
DWG- NO.
LS117
A
DO NOT SCALE DRAWING
ICAD
SCALE
2
FILE:
II
1
SHEET
1
OF
8
8
II
,
7
665
55
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine Pharmoceuticad ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
4
I
33
I
I
I
21)
I
T1
D
D
2.300
1.500e
.062
.250
-
U
C
2.000
1.103
.203 -.
175 THRU
FJ 0.290T.180
B
.312
-
.624
=
I
R.312
L0.120
(R.060)
U
T
SPECIICATI
ORREN^YNORNO.
DCRIPTO N
N0.
N
RED
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
4-40 TA P THRU
A
FRACTIONS
+
MATERIAL
DECIMALS
XX+ D1
.XXX+0OD5
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
APPROVALS
ANGLES
+ 1
Aluminum 5052
8
1
B71
I
1
--
66
I
1
T
'
Light Source
Grating Arm
RESP ENO
USED ON
APPLICATION
A
EC Hoorou March 2001
CHECKED
FINISH
NEXT ASSY
MIT Mechatronics Research Laboratory
DATE
DO NOT SCALE DRAWING
I
i
QUAL
SIZE IDWG.
NO.
SCALE
ICAD
LS118
A
ENG
j2
3
2
FILE:
SHEET
OF
7
8
I
6
5
1
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine Pharrmceuticol ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
8
7
6
4
4
4
4I
1
3
1
I
I
2
D
D
c
C
2.50
-111
-
-
.30
.E:
r
B
1.00 -
.50-
I
.25
.30
I
.
B
.9j0
F'3:=
L
.30
0.18 THRU
LJ 0.30-.16
PART OR
IDENTIFY NG NO.
ITEM
NO.
kRIAL
MA
SPECIFICATION
NOMENCLATURt
OR DESCRIPTION
UIT
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS DECIMALS
A
+
.XX+ .T
ANGLES
+1I
.xxx+.oo5
MATERIAL
Aluminum 5052
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
APPROVALS
MIT Mechatronics Research Laboratory
DATE
A
Eric Hoarau Feb 2001
Light Source
Grating Base Leg 1
CHECKED
RESP ENG
FINISH
NEXT ASSY
SIZE IDWG. NO.
USED ON
DO NOT SCALE DRAWING
APPLICATION
15
1I
//
1I
6
I
-
.L
-
44
LS1 19
A
QUAL ENG
I CAD
SCALE
3
j
L
41
FILE:
I
I SHEET
I
OF
1
8
5
4
1
3
I
2
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and ApIne Phtrmoceutica ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
~1.
clQ
C
-
*~.
c
3.30
US
.40-
0
-
.50
.25
.55
.64
R.I
0
-
1.00
.30
B
0.18 THRU
L_ 0.30T.16
PARTS
UNLESS OTHERWISE SPECIFIED
DMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACIONS
A
DECIMALS
+
XX+01
APPROVALS
I+
.xxx+mS
MATERIAL
Aluminum 5052
EiC Hoarau
UTT
REQD
LIST
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
ANGLES
+
MAEAL
SPECIPICATION
NomENCLAIURE
OR DESCRIPTION
PART OR
CDNTTFYING NO.
ITEM
NO.
MIT MeChatroniCs Research Laboratory
DATE
A
Feb 2001
Light Source
Grating base leg 2
CHECEED
RESP ENG
FINISH
NEXT ASSY
SIZE
USED ON
DO NOT SCALE DRAWING
APPUCATION
8
|
7
I
6
|
5
'
4
FOAL ENG
'T
IDWG.
NO.
CAD
SCALE
2
LS 120
SHEET
FILE:
|
1
OF
7
8
I THE INFORMATION
6
8
CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT ond Aopie Pharmoceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
1
~~
~
3
4
.
~
2
1I
D
D
0
2.000
c
c
.188
.125
-. 250
1.875
.094
I
.125
0.125
CL
.750
I
E
to
1.000
1.250
4-40 TAP ~ .500
L..I
B
j
IQ
ITEM
(IQ
PART
ORMATRIAL
ESENITFYING
NO.
B
SPECIFICATION
OR DESCRIPTION
NO.
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
A
+
DECIMALS
.XX+ .Ol
.XXX+0D5
ANGLES
+1I
MATERIAL
Aluminum 5052
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
Eric Hoarau
MIT Mechatronics Research Laboratory
DATE
A
Feb 2001
Light Source
Motor holder bracket
CHECKED
RE$P ENG
FINISR
NEXT ASSY
IS
8
1I
I
/
II
6I
i
I
SIZE
USED ON
APPLICATION
DO NOT SCALE DRAWING
4
4
I LWU. Nu.
LS 121
QUAL ENG
SCALE
CAD
FILE:
SHEET
OF
6
8
I
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MO ad
Akw Phamioceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMSSION OF IS PROHIBITED.
1
55
44
.L
33
1
2
I
0
D
D
1.654
0
C
0
rA
c
.063
4x 0.138
0R.156
0
Pi.
-Uf
0.313
6 .I
.610 .827
-
C
4-
2x 0.125
.827
B
0.900
.610
0
0
-. 827
-
.063
1.654
SECTION C-C
SCALE 1.5: 1
B
.188
|.610
C
SPECIFICATION
OR DES.RPIN
8=FNG NO.
.
RoD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
A
+
DECIMALS
TTT+ .01
ANGLES
+
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
DATE
EriC Hoarau
Feb 2001
MIT Mechatronics Research Laboratory
A
Light source
Motor holder
ZHECKED
MATERIAL
Aluminum 5052
TSP ENG
FINISH
NETASSEDON
8
I
I
6
I
APPUCATION
3
T
DO NOT SCALE DRAWING
4
UAL ENG
122
ALS
SCALE
ICAD
2I
2
FILE:
IREV,
SHEET
1
1
OF
8
6
7
4
5
I
3
I
i
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MT and Ampine Phcgmoceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMSSION IS PROHIBITED.
D
I
2
m
I
I
I
I
I
I
I
I
I
I
I
I
I
I
D
13
c
c
GI
CM
6.35
0-
5~ 13
P15
MB
This part is purchased
The small center hole is rebored to a
diameter of 5mm to fit on the motor shaft
B
ITEM
PART
OR
IDENTIFPING
NO.
MATERTAL
SPECIFICATION
NOMtNCLlUII
OR DESCRIPTION
NO.
B
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN Mffilmter
TOLERANCES ARE:
FRACTIONS
A
DECIMALS
+
X)+ .1
ANGLES
+ 1
.xx+.05
FINISH
8
1
7
1
6O
6
1
I
APLCTO
5R4
IDO
IDO
DATE
Eric Hoarau
Feb 2001
A
Light Source
Motor Flexible Coupling
I
MF~tNG
NOT
MIT Mechatronics Research Laboratory
RESP ENG
USED ON
APPLICATION
APPROVALS
CHECKED
MATERIAL
NEXT ASSY
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
SCALE
SCL
4
..
I
UU^"tNU
DRAWiNG
WIGI
1
SIZE IDWG-
3
NO.
LS123
A
|CAD
-SCALE
2
FILE:
i
SHEET
I
OF
C3 Purchased Parts
Deuterium Lamp
L6311-50, L6312-50
(
ARC POINT
30±11
(Costruction)
(External view)
052.0=0 5
LIGHT OUTPUT
SHMOW
xE
onAM
Led w"
CONNE
:AON
F;LAMENT.GNO. BLACK
AOE
I EO
Characteristics
1.0
I16.)
to
350
300 +/- 30
80
+/-0.3
0.05
2.5 +/- 0.25 4
20
I1.0+/- 0.1
1.8
4000
LOW'
1.0
185 to
350
300+/-30
80
+/-0.3
0.05
3.0+/-0.3
5
20
0 to 1
0 to 1.6
4000
L6301
0.5
185 to
400
300+/-30
80
+/-0.3
0.05
2.5+/-0.25
4
20
1.0+/-0.1
1.8
2000
16302
1.0
185 to
350
300+/-30
80
+/-0.3
0.05
2.5 +/-0.25
4
20
1.0+/-0.1
1.8
2000
1630
.5
5
18
1
to
to
400
300+/-30
80
+/-0.3
0.05
2.5+/-0.25
4
20
1.7+/-0.2
3.3
2000
130
0
185 to
350
300+/- 30
80
+/- 0.3
0.05
2.5+/-
0.25
4
20
17+/-0.2 3.3
2000
05
1
to
400
300+/- 30
80
+/- 0.3
0.05
3.0+/-
0.3
5
20
oto
0
0
18 to
185 to
350
350
180
300 +/- 30
300+/- 30
185
80
80
+/-0.3
+/- 0.3
0.05
30 +/- 03
5.
2.5to 6.
0.05
to0.3
10+/- 1
0.8
20
20
0.
18
400
300+/- 30
80
+/-0.3
0.05
10+/-
1
0.8
20
2.5 to60.6
______
____0.6_
80
80
+/-0.3
+/-0.3
0.05
0.05
10+/- 1
10+/-1
12
20
20
7 +/-0.5
.0
7.0+/-0.5
1
1.2
1
2000
2000
1.0
0
L6Vl
0.5
.32
1.0
to
400
185 to
184t030
350
400
300+/- 30
300+/-30
1
25to 6
0 to
1.6
3tt
to.
0 to
to
2000
2000
2000
2000
_____
to
400
300+/- 10
80
+/-0.3
0.05
12 to 15
0.5 to 0.55
20
0
0
2000
1350
300+/-10
80
+/-0.3
0.05
12 to 15
0.5 to 0.55 20
0
0
2000
*: The life end is defined as the time when the radiant intensity falls to 50% of its initial value or when the output
fluctuation exceeds 0.05%p-p.
D 2000 Hamamatsu.
147
Power Supply
CHARACTERISTICS
Anode output
parametCr
C444
Output current
Output voltage
80±20
160
80±10
160
mAdc
Vdc
Vdc Typ
600150
500
±0.05
Vpeak
% Max,
±0.05
±0.05
% Max.
%/h Max,
0.1
%p-p Max
10.05
Input fluctuation (± 10%)
Output fluctuation
0.05
±0.1
0.1
0.5A Fuse
Load fluctuation (normal operation range)
Drift
Ripple
Over-load protection
UAK,
300
Normal operation
No load
Trigger voltage
IC-4545
300
0.5A Fuse
-
Filament (heater) output
Output voltage for warm-up
10±1121
2.5±0.2
Warm-up time(Approx.)
Output voltage for operation
Vdc
20
20
s
7.0±0.5 / 3.5±0.5
17±0.2 / 1.010,1
0
Vdc
Input fluctuation (I10%)
±0.1
Over-load protection
% Max.
O1V side: 2A slow-blow
2.5V side: Fuse (5A)
Input voltage
Operating ambient temperature
Performance guaranteed temperature
Cooling method
Extemal dimensions (WX H XD)
Weight(Approx.)
100/118/230±10%
100/118/230±10%
0 to +40
+5 to +35
Oto+40
+5 to +35
Not required
200 X107X240
6.7
Not required
70x118X195 83x136x214
2
3.4
L6301. 16302, L6303,
16308, L6309. L6310,
L7174, L7291, L7292,
L7295, L7296, L7297,
L7304, L7305. L7306,
Applicable lamps
C1518
C4544
148
Fuse (1A)
L6304.
L6565.
L7293,
L7298,
L7307
L6307,
L6999,
7294,
L7303,
L631 1. L6311-50,
L631, L6311-50
L.312, L6312-50
C4545
Vac
mm
kg
Stepper Motor 4SQ-120BA34S (for light source and transportation system)
Not available for sale in Europe
Series 4SQ Stepper Motors 1.80
Dimensions: mm/in
-
42'2
07±2
I911111
k2714
401~20
_6'
20.16o
144
so
--
--
--
-
40
71
.35
PULL
00
.2
-
2r
"o
-
5
Wiring
Diagram
7
--
0
00oo
-
100
-
200
4-
-
200
o -o
-
400 500
24
1
-o
-
WO0
1042
07
-
70k) Boo6
Specifications
00
_
1_
Partfuber
4
1A0DAW
NOTE: Refer to page 7
12
Resi~per,',Wpdino .'
5
And,,portWhdihginnH
Roto
Det'ntToq ue, rnr
-
Catalog P/N Construction
26
6M2
Mgfontfilatta
Example 1. Single,Shaft Extension
X10"
ent1.9
z
8.5/01.2
Step~~gla1
4_-
060
BA
I[
.8t
4L:Single Shaft Extension
L34 mm/ .3 Length
si5%
X-
A
3
MMP~r;RM200
4/8.8
w 4,
/
- Mag~iti~sait
g55
'Operating,
P
Aluminum End Bells
6 Volts
Basic Motor
kg7i5/17.6
0C
-20*C to +500C
-20'C to +60*C
bal, Double Shielded
r'glypsq
50
Example 2. Double Shaft Extension
4
- 1120
F
Steel
-- 12 Volts
Basic Motor
For Information or to place an order In North America: 1 (203) 271-444
M C AT
ONC
M
T-3
NOTE uIjes othwee wncaed al vaus ahown are tyocul Other wndngs aileye
on spcoal order Conani Thomson Arpax For avWiaSy of motoMr
30a' step nge.
'Meased
rWl2 pas energad
"MeaedttOmom mtmlOnnorlgpm p 1e4sace
PA
BFA
-eohms
500 Vac for 60 S
l~e tng/az195/7
26
for unipolar switching sequence
Double Shaft Extension
mm/1.3 Length
End Bells
4
Asia: (65) 7474-888
The 26secat in Oft pubcatio we boe
to be auTa and reshNlC
However, iti the sPQWlb6%y of the
produCt useWI to
tMo. the wtataity ot Thomso products torta spealc spllceon, Winl e tfective products cl
be r"Wc ihu
fpn~
. n tiedty is =vned beOd udreplalanet.
ex
~g
149
Electronic Shutter: Prontor Magnetic 016 from Schneider Optics
"jy
Z"eae-GJUpp
Size
PRONTOR magnetic
Toille
Bestoll - Nf.
F010
Grdse* 0
0
PRONTOR magnetic
magnetic
64V""e IPRONTOR
0
I
I
Order No.
No de conmonde
Selichtungsz eiten:
Exposure times :
32s 16s 8s 4s 2a is
Blende :
Offnung 23 0
Gr&Pte
Kleinste bffnung 1,5 0
1/2s 1/4s 1/8s 1/15s 1/30s
Diaphragm
Maximum aperture 23 0
Minimum aperture 1,5 0
Gesamthbhe : 2Dm+O,o 3
Abstand der ObjektivrohrstirnflAchen
Gehiusedurchmesser :
61mm
Total height : 20vr+o,0 3
Distance between the front
and the rear faces of the
lens tube
Housing diameter : 61. tw
Temps do
pose:
1/60s.
Diaphragme
Ouverture max. 23 0
Ouverture min. 1,5 0
Hauteur totals : 2knm+O,o3
Distance entrv les faces
du tube porte-objectif
Diamdtra du boltier : 61 nn
Fassungsgewinde
vorn und hinten : M29,5x0,5
Filetage avant at
Front and rear lens
mounting thread: H 29,5x0,5 erriire:
M 29,5 x 0,5
Anschraubgewinde: M32,5x0,5
Thread of retaining ring:
M 32,5 x 0,5
Shortest possible
exposure time . 1160s
Filetage de Ia begue de
fixation: M 32,5 x 0,5
Temps de pose le
i/60s
plus court :
Magnetic drive (long life)
remote speed control with
power pack, electronically
controlled shutter speeds,
manually controlled aperture continuously
adjustable.
Fonctionnement magnetique
(grande longevite),
commande a distance par
l'intermedisire d'un
dispositif de comwande se
branchant sur Is secteur,
vitesses d'obturation
pilotmes electriquement,
diaphragme reglable
manuellement' sans crans
kZrzeste mgliche
Belichtungszeit
1/60s
Magnetischer Antrieb
(hohe Lebensdauer), Fernbedienung .mittels Steuergerbt mit NetzanschluS,
alektronisch- gesteuerte
BelichtungazeIten, manuell
einstellbare stufenlose
Blende.
d'imnmobilisation.
FOr MaBe und Daten sind nur
Angaben der Originalzeichnung 1016 602 verbindlich.
For dimensions and other datalSeules lea dinerisions et les
original factory drw.iing
donnees du dessin orginal d'
--'I
---^1.- 4nA
in
1 016 602 is avalilabh.
150
Appendix D
The Excision Device
Dl Assembly Drawings
151
8
,
8
7
5
3
4
.15
THE INFORMATION CONTAINED IN THIS DRAiNG IS THE SOLE PROPERTY OF
MIT and Adpln Phczygcetild ANY REPRODUICTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
1
2
2
1
2
1
0
D
15
D
00
00
c
7
1Cutting blades (glued to cutting tip with epoxy)
'
LA
A
2 Cutting tip
14
3 Cutting tip holder
bolte to slide by M2.6, 3mm in front, 6mm in back)
4 Positioning pins (1/16" diameter, 5/16" length
press fitted to cutting tip holder)
5 Magnet (glued to cutting tip with epoxy
and #4 nut glued to cutting tip holder)
6 Stopper blocks
(bolted with a 0-80, 5/16" cap screw)
7 Spring
8 Motor base
(bolted to slide by 2x M2.6, 6mm with #4 washer)
9 Stopping pins (1/1 6"x5/16")
(press fitted 3/32"deep)
10 Slide (BSP 10-25 SL)
(bolted to motor base by 2x M2.6, 4mm cap screws)
11 Limit switches
(bolted with 2x 0-80,1/4" cap screws)
12 Digital linear actuator (L92121 -P2)
(bolted to its holder by 2x 6-32,1/4" screws)
13 Main frame
14 Slide (BSP 10-45 SL)
(bolted to main base by 2x M2.6, 4mm cap screws
15 Motor Shaft (cut to 3" total length)
(hold to the shaft plate by #4 nut and washer)
0-
11
B
3
IEM
UNLESS O
6
RE
SE
DTOLERNCEOS ARE:I
FRACIONS
DECIMALS
x+
+
-XX+Dl5
MATERIAL
iPIE
D
~rl
NEXT ASSY
5
CAD GENERATED DRAWING.
DO NOT MANUALLT UPDATE
APPROVALS
ANGLES
.1 + 1
SPECIFICATON
REOD
RED
Eric Hoarau
Aluminum 5Ct5
-
Excising Tool
Assembly
tNu
SIZE
QUAL ENG
DO NOT SCALE DRAWING
4
A
Feb 2001
RESP EINGD
M-A
USEDON
MIT Mechatronics Research Laboratory
DATE
CHECKED
FINEH
R
APPUCATION
7/
MATERIAL
NOtCA~&|
OR DESCRIPTION
PARTS LIST
16 Shaft plate
(bolted the main frame by a 4-40,1/4" and a #4 steel washer
a #4 plastic washer is placed between the plate and frame
OR
PART
IDENTIFY NG NO.
N.1
DWT.
1
35
3
1I
NO.
CAD FILE:
SCALE
2
ETOOI
I
I
SHEET
I
OF
D2 Parts Drawings
153
8
7
6
5
7
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
8
MT and Ahpie Pharrnaceutical ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMSSION IS PROHIBITED.
7ID
7
17.50
-25.40-'
4
S4
6
4
I
S3
1
2
2
3
8
-1
0 1.50
00
S
0 P
2x 4-40 TAP TRHU
D
2.5
00
15.50
12.50
70
-- 3
p.'
c
11
4-40 TAP THRU-
50
-
05 THRU
31
W0o
B
3.80
48.504x
F7.50
81
1
PRT OR
E NTIFYINORNO.
ITEM
'NO
Noe
OR
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN Milneter
TOLERANCES ARE:
A
2x 03THRU
LJ 0 5W2.60
FRACTIONS
A
-
I
66
-
ANGLES
X+E.A+T
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
I
RD
MIT Mechotronics Research Laboratory
DATE
APPROVALS
A
Eric Hoarau Feb 2001
Excising Tool
Main Frame
MATERIAL
CESP ENG
Aluminum 5052
5
NEXT ASSY
7
TRAI
SPECCATION
CMECKED
-
5
SE
USED ON
APPLICATION
~
DECIMALS
+
T
35
[-
AMA
RPTION
PARTS LIST
41
0
I
.
7.501
8
8
35
0-80TAP THRU
DO NOT SCALE DRAWING
4
4
F
1
REV.
DWG. NO.
ET101
UAL ENG
3
3
i
SCALE
2
2
ICAD
PILE:
I
SHEET
1
1
1
OF
8
6
7
5
.L
3
44
.
35
D
2x
2 THRU
-
1I
2
2
THE IHFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
WffTnd AE RMSgocuicd ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
0
5
00
D
0
0
0
20
15
0)
7.50
01.50W2.50
45
16.5&-
c
3.50
-
10
-7
I-
2x 6-32 TAP THRU
'-
17.50
44.50
IL
1750
22
015 THRU
-0
-B
B
3.50
30
T_
70.50
H2 e*
I7
--
2x 03THRU
J 0 4.75W 1.56
20 -e
5-
ITM
13.50
30
1
0
A
0
13.18
-T
.,-2x 0-80 TAP THRU
-7
O
NO
OR
wtN;AU
O
FYNG NO.
FRACTIONS
DECIMALS
+
'X+'l.
ANGLES
1
MIT Mechatronics Research Laboratory
Eric Hoarau Feb 2001
Excising Tool
Small Frame
CHECKED
0-80 TAP THRU
FINISH
17.50
NEXT ASSY
I
I
6
1
USED ON
APPLICATION
5
' DO NOT SCALE
4
RED
DATE
Aluminum 5052
-
T
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
SPECIFICATON
DESCRIPTION
PARTS
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN MilliRmeter
TOLERANCES ARE:
MAEIL
MATERIAL
12.50
8
PART
7
G.
SZEIDW1
SCALE
1
1O
IRlv.
AET102
:DUAL ENG
DRAWING
3
ICAD
FILE:
I SHEET
I
OF
8
II
THE
TH
7
6
DRAWII
ITHE SOE PROPE
O
REPRODUCTION IN PART OR WHOLE WITHOUT
CONTAINED IN THIS DRAWiNG IS THE SOLE PROPERTY OP
8
INFORMATION
IFRAIO
CNANED IN THI
MIT aNd A~se Ptamocwuical ANY
5
1
4
7
1
4
6
3
I
2
1
2
WRITTEN PERMISSION IS PROHIBITED.I
03 THRU
Q o
D
D
0
107
03 THRU
52.50
0_5T
C
10 --
5
SECTION A-A
C
0 1THRU
17
10-
-
2
.5
2
05.50W2
1.50~31.60
2x
7
4-
20
~.
/
13 --
A
A
B
B
3.70
t~~
0
6.50 -
a ~
-17
12
L J
0-80 TAP THRU
5 *
T0.50
1.63
A
ITE
PART
r-
-
6.60
-
OR,
PCIEIA
PARTS
t10
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN Milmeer
TOLERANCES ARE:
3.25
0
FRACTIONS
+
DECIMALS
.X4-.'
.XX+.05
ANGLES
+1I
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
DATE
Eric Hoarau
Feb 2001
MIT Mechatronics Research Laboratory
A
Excising Tool
Cutting Tip Holder
CHECKED
Aluminum 5052
RESP ENG
FINISH
NEXT ASSY
1
7
1
6
1
5
SIZE
USED ON
DO NOT SCALE DRAWING
APPUCATION
9
4
*1
R
__________________
MATERIAL
17
8
SPECFICATON
ORDEIPTION
"FYING NO.
"N.
DWG.
ET1Q3
NO.
:UAL ENG
CAD
SCALE
3
2
SHEET
FILE:
I1
OF
T
II
8
1
7
3
1
4
jr
5
1
6
1
2
1
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Ampine Phrmnceuticad ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
276-
4S
D
0.039 THRU
0.217T.059
.512
0.071 T.276
C
.260
.079
A
t
.669
.413
I
c
0-
SECTION A-A
.500 -ft
B
B
.440
ITE
PART
OR
f
AE.A
LUR
N
SPECIIC
DESCRIPTION
TIFYING N.
NO
REOD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
A
+
MATERIAL
-
0
//
I1
6
0
I1
MIT Mechatronics Research Laboratory
DATE
A
DRAWN
Eric Hoarau Feb 2001
Excising Tool
CHECKED
RESPEND
DO NOT SCALE DRAWING
rUAL ENG
Cutting Tip Base
SIZE
.L
5
+1I
APPROVALS
USED ON
APPLCATION
I1
.XX+ DT
.XXX+.005
ANGLES
Aluminum 5052
INISH
NEXT ASSY
DECIMALS
CADGENERATED DRAWING,
DO NOT MANUALLY UPDATE
4
4
-~-
DWO.
NO.
ET104
A
SCALE
3
13
I
IS
CAD FILE:
I
I
OF
I
8
6
6
8
THE INFORMATION
CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine PharmaceutiCk
ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
1
5
1
4
&
5
3
1
1
22
1
I
I R 1.40 W 1.50
D
A
A
3
RO.50
3.25
os
1.63
4L '
01.35 THRU
c
00
D
c
SECTION A-A
SCALE 7: 1
il
7
00
6.25
Pil
B
B
/
/
9.50
ITE
PART NOR.
DTFYENOSNO.
NO.
OR
-ATA
SPECI ICATION
RIPTION
D
REaD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN Milmeter
TOLERANCES ARE:
FRACTIONS
2 Part needed
A
+
DECIMALS
X+ .T
.X)(+.05
ANGLES
+ 1
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
DATE
Eric Hoarau
Feb 2001
CHECKED
MATERIAL
Aluminum 5052
FINISH
NEXT ASSY
d
1
1
6
1
5
j DO NOT SCALE DRAWING
T
A
Excising Tool
Stopper block
-
USED ON
APPUCATION
MIT Mechatronics Research Laboratory
APPROVALS
.
oUAL ENG
3
ELE~
SCALE
SCALE
IICAD
CAD FiLE:
____I
Z
ET105
SHEET
1
",:V,
I
OF
1
8
5
L
4
3
1
I
2
1
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Alpine PharmoceuticaI ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
Q)
0.33
21
-C
11
c
'o -5
RI-N
1
.I
F
5
B
V
R
J
2x 03.20 THRU
I
TEM
"O
OR
PART OR
IENTIFYING
NO.
NO.
M
A=
:ALTER
SPECIFICA
DESCRIPTiON
ON
REQD
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN Mifimter
TOLERANCES ARE:
This component is made
from a spring metal sheet
A
FRACTIONS
DECIMALS
.X+ .
+
ANGLES
+1I
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
MIT Mechatronics Research Laboratory
DATE
APPROVALS
Eric Hoarau
A
Feb 2001
Excising Tool
Flexible Shaft Plate
CHECKED
MATERIAL
spring steel
NEXT ASSY
S/
I
1
I
6
I
SIZE DWG.
A
USED ON
ICATION
/6
I
RSPENG
I
DO NOT
SCALE DRAWING
4
4
PUAL ENG
3
j
SCALE
NO.
ET106
[CAD FILE:
I SHEET
IOf
D3 Purchased Parts
Digital Linear Actuator L92121-P2
14.86 ±025
21 08
2830
-
THD
10
56
t051
.02
06
Typical Linear Pull-in Force vs. Linear Rate at 20'C
48
WITHIN
iLKOo SH1OERj
"692
MA)
Nc-2A.
#4-40
4
F
11
F-1
*002
1
.5,0
25,9
A
2
42.93
005)
.0(0
5-8
1168
0.0
-
'.25
-6
-4---45*00
.062
366
E.00
Specifications
##i ipNinn
mna axim w
I
T el-
4
'Il-oz2
1921112*
12
0.5" (12.7mm)
5
12
5
2
wji2i4
KW
rim'UI
300
1 0
5 O
0.0
50
400
14
,
01
0.8
0.3
0.4
45
500
.8
0.8
0.1,
0.5
6
$TEN/SEC
20
IN/SE
0.98
.001" (.025mm)
minimom
kgldlngl~or
n
(UnenerglzedY
___P _
Max'R
B
psial
60 oz (1668N)
45 oz (12.5N)
0.5" (12.7mm)
,002" (05mm)
28 oz (7.2-3N)
1.5 oz. (42.5 91)
to 70"*
C':-2O9C
t-'
40 oz (11.1 3N).
-400C to
Colt Data
12
5
2421-02
250
200
15C
9C 100
006
ois
j'
4 0.
01 2
0.10
0.25
0 02
' 00 0 1
1,875' (47.6m
5
P
'42'
'((11 002'i
'V 2 1
+.03
Fortm
Per
i
K /2
0
4.5 WattS
tr
1.87T,
85?C
-Pi (5Vdo)
P (2VC)
ISO
641I
(476m)
,12
5
0.50 (12.7mm)
A*"21-Pt
12
L9214%P2
5 1.875" (47.6mnm)
12__
.004" (.1Omm)
I oz (195N
1602 (445N)
Nate: Shaft Options Series K92100
Add Suffix-S1 fbr #4-40 NC-2A Thraded Tip
Add Suffix-S2 for #2-56 NC-2A Threaded Tip
Standard Switching Seqtuence for Linear Actuators - Unipolar Dive 5Vda and 12Vdc
cg
G'
Q4
f%
QFF
i
F
0
0FF
OFF
1
Lead Wire ColorrCodes
ORN
92
FF
1 #
OFF
9
U~nipotar Drive
Note: Chart sequence,
QI QZ
(See hart
QGQ for proper
crtIor Gd)
BLK
Y BkU
-
.,FOF
OFF
from top of dhart to bottom.
For VtWr Thrust, uSe
SltiA§ fm .' tt~
chart to,top.
160
E
LEADS
02
7620
7TE38
28 AWG PVC
013
44 *.005
(2) HOLES
K
ST
I.
4
038
11,30
4
-
____
-
- -
0t
3
EL9
(9
yo
c 0p1
BLU
---
/7fff~fj~E
ORN
_
WNT
RED
Ei
RE0
WI-T-1~
RED
-RN - - -
(0
D4 The protean 2-D Spot Cutter by Bio-Rad
With jpotQn software, spot selection is
simple and flexible.
Specifications
Accuracy
* 0.1 mm in 10 consecutive cuts.
> 95% spot pickup on first cut.
0.01 mm increments for cutting head movement.
Cuts 96 spots in 20 minutes.
Sample Output
Reduces levels of keratin contamination, as detected in mass
spectrometry.
No protein carryover detectable in mass spectrometry.
Hardware
Excision Tip
Internal diameter 1.0 mm; outer diameter 1.5 mm.
Cuts acrylamide gels and PVDF membranes.
Dimensions (without computer)
Size: 50 x 52 x 50 cm (W x H x D);
weight: 30 kg (66 lb)
Lighting
White light under cutting stage and microtiter plate.
Flash in camera for overhead lighting.
Regulatory
CE, EN61010-1
Maximum Cutting Area
11.8 x 8.8 cm (W x H)
Accessories
11.8 x 8.8 cm (W x H) large and 9.5 x 7 (W x H) cm mini gel sizers
and frames, cutting mats, cutting tips, starter kit
Imaging System
Picture resolution: 1280 x 960 pixels (W x H)
Operation
Operating temperature: 10-30 *C.
Operating humidity range: 0-95% RH non-condensing.
Minimum Computer
Requirements (computer
not included)
PC - 300 MX (Pentium® II or equivalent), Windows® 95/98/NT,
64 MB RAM, PS2 mouse, 2 GB hard drive, 24 speed CD ROM
drive, 15" XVGA monitor, 24-bit color, 1024 x 768 resolution
PowerPointand Windows are registeredtrademarksof Microsoft Corporation.Pentium is a trademark
ofIntel Corporation.
All content copyright Bio-Rad Laboratories Inc. @ 2001.
All trademarks and registered trademarks are the property of their respective companies.
161
Appendix E
The Excision Device Transportation System
162
El Derivations for the transportation system
E1.1
System Torque Calculation
T
0
00
Figure El: Schematic of the rack and pinion system
The total torque required by the motor is
+Tfriction
Total :Trotational +Tinear
acceleration
acceleration
(E.1)
Totational is the torque required to rotate the rotor, the shaft, and the pinion.
acceleration
a
Trotational
acceleration
=Jrotor
+ pinion
=rotor
+ pinion
a
R
(E.2)
where
Jrotor
is the moment of inertia,
+ pinion
a
R
is the linear acceleration,
is the pitch radius of the pinion gear.
The torque required to accelerate the carriage and its components is:
T
inear
acceleration
(E.3)
= maR
where m is the mass of the moving assembly.
M=mcutter + mmotor +mframe.
(E.4)
The frictional torque, Tfriction, is the torque required to overcome the frictional forces, Ffriction,
between the slide-rail system and between the rack-pinion system.
(E.5)
Tfriction =FfrictionR
The total required torque is therefore:
163
a
;ota
J,,
-+maRr+oFrfricton
+ pinion R
(E.6)
The values for the different variables in the designed transportation assembly are:
mframe
= 160 g
= 197g
= 206g
Jrotor
= 1.9x10 3 g. m
mcutter
mmotor
2
+ pinion
R
= 0.016 m
a
= 0.15m/s 2
Fficion
= 1.37N to 1.77N
The acceleration was obtained from the specification of reaching the cruising speed of .15m/s in
1 second. The frictional force for the entire assembly was determined experimentally by using a
pulley system and calibrated weights.
It is expressed as a range because it was found to be
different at different locations. The stepper motor therefore needs to provide a torque equal to:
=0.02+1.35+21.92= 23.29mN -m
Tfxtota =0.02+1.35+28.32= 29.69mN -m
TWntota
The maximum operating range of the proposed system is shown on top of the motor torque
performance in Figure E.2. The selected motor can be used at up to 300-pulses-per-second with
a safety factor.
Operating
range
45
-Or
-
A8.
5-0
4.2
PULLOUT
0
--
-
0
10.
-
0
100
1
121
-is
20
300
400 50
W0
SPUD (P58)
7M)
M0
M0000
Figure E2: Torque curves for the 4SQ-120BA34S stepper motor
(Courtesy of Thomson Airpax Mechatronics)
164
E1.2
Calculation of System Resolution and Speed
The linear step size, A,, is calculated from the stepper motor step angle, A, and the pitch radius,
Rp, of the pinion gear.
Al =
R
360/A,
(E.7)
The selected stepper motor has a 1.80 step angle. The linear step size is therefore 0.5026mm.
The speed, V, is calculated from the step size and the pulse frequency, fi.
V=2;rRp 3fM
(E.8)
As seen in the previous section, the stepper motor maximum operating pulse frequency is 300Hz
for the proposed application. At 300pps, the system has a linear velocity of 150.8mm/s. The
transportation system speed can be estimated to be half of the pulse frequency.
E1.3
Calculation of Rack and Pinion Specifications
During motion, it is safe to assume that one tooth is supporting the load. The gear tooth strength
is found from the Lewis bending strength equation:
SFY
(E.9)
W =S
where
Wt
S
F
Y
is the maximum transmitted load,
is the maximum bending tooth stress and is equal to one-third the tensile strength,
is the face width of the gear
is the Lewis factor,
Dpith is the diametral pitch
Both the gear and racks are made of 303-stainless steel, so the tensile strength is 90,000psi. The
pinion gear has a face width of 0. 104inch, which is smaller than the one of the racks; therefore, it
will be used. The Lewis factor is equal to 0.421 since we are using a 60tooth gear. The racks
and pinion have a 48-diametral pitch. The maximum transmitted load is therefore 27.4lbs or
122N.
The linear pitch, Lpih,, is the distance between two adjacent teeth.
Lpitchj
D
=
0.0654inch =1.7mm
pitch
165
(E.10)
E2 Parts Drawings
166
8
I
6
5
3
4
L
11
22
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
WTand Alpne Phamoceutica
ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
D
D
c
3
rA
0-1
WI
A
1 Main Frame
2 Accuglide Rail
Bolted to main frame by 25x 8-32,7/16"
socket cap screws
3 Accuglide slider
4 Racks
Each rack is bolted to main frame by
4x 6-32,1/2" button head cap screws
5 Stepper motor, model 4SQ-1 20BA34S
Attached to carriage frame #2 by 2x M3 nuts
6 Anti-backlash pinion gear, model AP48C-60
Hole is rebored to fit the 5mm diameter motor shaft
7 Carriage frame #1
Bolted to slider by 4x M4,6mm cap screws
8 Carriage frame #2
Bolted to carriage frame #1 by 3x 6-32,5/8"
cap screws and spring washer
9 Excision device
Bolted to carriage frame #2 by 2x 4-40,1/4"
cap screws and washers
INEXT ASSYJ
2
a
0
I
66
I4
b
B
5
9
PARTS
UNLESS OTHERWISE SPECIFIED.
DIWNSIONS ARE IN Milrireter
TOLERANCES ARE:
FRACTIONS
+
DECIMALS
X+ .T
.XX+5
ANGLES
+
1
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
MIT Mechatronics Research Laboratory
DATE
ErEc Hoarau Feb 2001
Assembly Drawing
RESP ENG
USED
-
ON
SIZE
DO NOT
A
Transportation System
CHECKED
MATERIAL
FINISH
APPLCATION
I
.
7
SCALE
4
DRAWING
DWG. NO.
TSOO 1
A
EN I
j
j
I
1
SCALE
2
2
CAI
IE
I
1
I
SHEET
1
1
REV.
Of
6-
8A
4
55-
211
3
32
1-II
I
KV and A4,e PInc~eufcd ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
25x 8-32 TAP THRU
D
D
.787
1.575
-C
.787
DETAIL C
SCALE 1 : 3
00
2.559
-0
39.370
C
0
0
0
0
0
0
0
C
0
0
0
25x 8-32 tapped holes are needed
~J7.3~ V
W
0'
M
-
z
x
B
0 A
0
B
0
0
00
0
.250
2.309+-
2x 0.343THRU
PART.
ID
NuS
OR
NG NOR
.374
UNLESS OTHERWISE SPECIFIED
DMENSIONS ARE IN INCHES
.787
.197
TOLERANCES ARE:
FRACTIONS
+
.315
A
.472
4x 6-32 TAP THRU
2x 0.343 THRU
DETAIL B
SCALE 1: 2
1
6
ANGLES
+ 1
MATERIAL
Aluminum 5052
DETAIL A
SCALE 1: 2
1
DECIMALS
.XX+ .01
.XXX+.WS5
NEXT ASSY
NECIFICATION
LIST
CAD GENERATED DRAWING
GO NOT MANUALLY UPDATE
MIT Mechatronics Research Laboratory
APPROVALS
DATE
Eric Hoarau
Feb 2001
A
Transportation System
Main Frame
CHECKED
RESP NG
SZE DWG. NO.
USED ON
^P"^"IN
1
1
A
DESCRIPTION
PARTS
s
.250
1.000
4 sets of 4x 6-32 tapped holes are needed
I ITEMI
NO
.787
B
1.476
2.8754
8.894
- .750
1.727
(00
0
2.875
2.875
H250
DO NOT SCALE DRAWING
A-
QUAL ENG
SCALE
A
1
CAD
FILE:
SHEET
1
OF
8
I
7
6
8
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and AlpIne Pharmceutick
ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
5
I
4
L
3
3
I
1I
2
6
I
D
0
D
=9
SECTION A-A
-. 250
T
0
-I
0
C
~1
C
3x 6-32 THRU
CLEARANCE HOLES-
9
--
.945 :
.945
.2.236
A
r .125
Cd,
0- T
0
)0
A
1.102
TI,
TI,
B
4x 0.177THRU
LJ 0.315-.157
1.575
B
1.969
L.1
.
9
9
9
-
1.181
PART
0TEM
O
.591
-
NTIFY
ARr
QT
SPECIFICATION
REOD
OEI
IOR
ING NO.
OR
DRPTON
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
2.362
FRACTIONS
A
+
DECIMALS
-XX+
01
ANGLES
+
1
.xxx+.0oS
CAD GENERATED DRAWING,
DO NOT MANUALLY UPDATE
APPROVALS
MIT Mechatronics Research Laboratory
DATE
A
TncNHoarau Feb 2001
Transportation System
Carriage Frame #1
CHECKED
MATERIAL
Aluminum 5052
FINISH
NEXT ASSY
DO NOT SCALE DRAWING
APPLICATION
0
v
I
-.
/
I1
0
0
I1
3.-
0
SZE
USED ON
-
TF
I
I
i
QUAL ENO
3
15
SCALE
I CAD
IRV.
T102
WG NO.
FILE:
I
1
I SHEET
I
1
OF
8
I
7
6
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and AIIne Phcrmceutilcd ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
1,
5
.
4
1
3
I
3
2
2
I
1
I
2.362
.945 t
D
.945
- .236
.125
D
0
3x 0.138W.787
.250
K
1.181 -
610 -
C
0.90 THRU
.300
.6 0
0
C
19
L
.300
1.220
4-
1.220
2x 0.130 TH RU
-
L J .375T.118
5.906
B
B
2.750
-
ITE
PART, OR
RIENTI NG NO.
NO
MAIA.
O
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
.375
FRACTIONS
A
+.
1.181
0.125 THRU
LJ 0.375T.188
DECIMALS
.XX+fll
.XXX+.IS5
ANGLES
+1I
Eic Hoarau
O
MIT Mechatronics Research Laboratory
DATE
A
Feb 2001
Transportation System
Carriage Frame #2
CHECKED
MATERIAL.
Aluminum 5052
71
LIST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
SPECIFICATON
RIPiON
PARTS
RESP ENG
FINISH
NEXT ASSY
IS
ts
II
//
I1
66
I
1
SIZE DING. NO.
USED ON
APPLICATION
DO NOT SCALE DRAWING
QUAL
ENG
A
SCALE
2
2
CAD FILE:
TS 103
I
I
REV.
ISHEET
1
OF
7
8
II
6
THE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
MIT and Apkw PhamaceuticdA ANY REPRODUCTION IN PART OR WHOLE WITHOUT
WRITTEN PERMISSION IS PROHIBITED.
1
8~
5
A,
4
1
3
1
1
2
1
D
D
c
8.894
-
0v
*
e
-
0q
o-1
B
4 parts needed
Model AG-1 purchased from Pic Design Inc,
0.053" were removed from each end so that the racks
could be stacked together
ITEM
PART
MAIERAL
ORNOMENCrLAURE
SPEC
I ICAT
OR DESCRIPTION
IEN TIFYING NO.
NO
SUIT
D
N
PARTS LIST
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
A
FRACTIONS
DECIMALS
+
.XX+ Dl
XX+W
ANGLES
+1
* 1
I
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
MRAWRN
Eric Hoarau
MIT Mechatronics Research Laboratory
DATE
A
Feb 2001
Transportation System
Rack, 48 pitch
CHECKED
MATEI AL
416 Stainless steel
FINISH
NEXT ASSY
8a
I1
7/
I1
66
I1
USED
APPUCATION
55
SIE 1DWGONO,
ON
~
DO NOT SCALE DRAWING
4
~
1
TS:4.
TS 04 I
A
rUAL EG
3
:3
I
1
SCALE
2
2
ICAD
FILE:
I
1
SHEET
1
1
OF
Appendix F
The Cutting Tip Changing Station
Fl Part Drawings
172
8
.
7
I I THE INFORMATION CONTAINED IN TIS DRAWNG IS THE SOLE PROPERTY Or
1I
8
6
57
MIT Ord Alpne Phamoceu:ic. ANY REPRODUCTION IN PART OR WHOLE WMHOfT
T-E WRIfTEN PERMISSION IS PROHIBITED.
4
22
-
I
11
I
I
B
D
B
B
6
D
B
08
910
0
00
0
01
0
C
VS
-0
C
1 Base plate
2 Tool Centering end piece
(bolted to base plate with 2x 8-32,7/1611
button head screws and washers)
sliding arm
B 34 Delrin
Shaft holder
(botted to base plate with 2x 8-32,7/16"
button head screws and washers)
B
5 Shaft
(attached to its holder by 2x 4-40 set screws)
6 Solenoid frame
(attached to base by 2x 6-32, 1/4" cap screws
and # 6 washers)
7 sliding arm bracket
(attached to sliding arm by 2x 4-40,3/16
ITEM
No
button head screws, and attached to solenok:1
arm by
PART OR
IDENTTFYING NO.
1x 4-40,1/4" screw)
UNESS OTHERWISE SPECIRED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACIONS DECIMALS ANGLES
8 Solenoid arm
A 9 Set screws to grab cutting tools
10 Front springs
+
11 Back springs
=+ M
APPROVAL$
USED
I
(
I
0
I
0
-
I
DO NOT SCALE DRAWING
_________________
I
'4
MIT Mechatronics Research Laboratory
Cutting Tip Changing Station:
A
Assembly Drawing
E
ON
APPUcATION
P'CNAT
EW_
__________________
DATE
URAWN
Feb 2001
Eric Hoarau
Abe Schnelde
+1
MATERIAL
U
UST
CAD GENERATRD DRAWING
DO NOT MANUALLY UIPDATE
FNISH
NEXT ASSY
z
OR DESCRIPTION
___________________PAMS
QJAL ENG
I
3
J...........,......~....I
SIDNN
SCAE
CAD FIL
SCALE ICAD FILE:
S00 1
REV.SPRE
|~hl
I
I
I
I
O
8
7
557
6
TH INFORMATION CONJTAINED IN THS DRAWING IS THE SOLE PROPEMrY OF
|T "d AO-e Rxffm caL ANY REPRODUCTION IN PAR OR V*POLE WITHOUT
TiE WRITEN PERMSSION IS PROHBITED.
8
4
4
6
33
I
1
22
7.00-
-1
D
.64 -
3.18--
D
SECTION B-B
SCALE 1: 2
2x 0.15THRU
2x 0.175THRU
LJ 0.45T. 15
Li 0.39W.131 \
C
C
1.50
6.00
Oil
4
.40
B
B
*
L~J]
0
1.28
.15 THRU .73
r--i r.28
L--] 1_--J
2.70
.55
L]--
1.69
A
5x
LT.03'
A
45
-
T
*
_
B
x
1.03
1111111.
L
.68
-t
B
2x 0.18 THRU
LiJ 0.45T.15
69
S1.40-aw
-2.32
PART
I
TM
NO
ORNMECA
NO.
IDENTFYNG
Z
Z
Z
3T---rr/ I I
A
H Al
i F Tr-M/IT---Fr
Z
LR*IESS OTHEWGSE SECFIED
DMENSNS ARE IN INCH
TOLERANCES ARE:
Z A
FRACTONS
+
SECTION A-A
SCALE 1: 2
DECMALS
XX+
ANGLES
a
+
MATERIAL
Clear plexigloss
NEXTASSY
E
APPLICATION
I
/
I
0
I
O
~PARTS
LIST
CAD GENEATEA DAING,
S0 NOT MANUALLY UPDATE
APPROVALS
DATE
Eric Hoarou
Abe Schne.
Feb 2001
1
SPEMFCATION
CeSCAPO
0,
____________________
WES);
__________________
MIT Mechotronics Research Laboratory
A
utting Tip Changing Station:
3ase
INSH
DO NOT SCALE
RAWNG
03101
A
SCALE
2
2
ICAC
I
i
ILE
[bIItI
|
LI
8
55
6
,
CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
THE INFORMATION
MT nd pre Phamoeutlcd. ANY REPRODUCTON IN PART OR WHOLE WITHOUT
4
DETAIL A
FSCALE 2 :
1
2
0
8-32 TAP THRU
.
D
33
.33
1.9
D
.04
.38 *
.47
.67
.25
A
.4
C
C
2x 0. 13 THRU
B
I
.50
.38
10
I
I
I
I
B
i
6.00
PAWE OR
WENTI"NG NO.
ITM
No
-
.3 8
-I
W*tS(LJf
OR DESCRPON
S
A
ON
PARTS UST
UNLESS OTHERWISE SPECIRED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS DECIMALS ANGLES
A
+
+.XX
+
MATRIAL
Aluminum 5052
RNISH
NEXT ASSY
15
b
I
I
/
II
0
I
I
U
APPUCATION
CAD GENERATED DRAWNNG,
DO NOT MANUALLY UPDATE
APPRIOVALS
Eric Hoarau Feb 2001
Abe Schnelc er
Cuffing Tip Changing Station:
Centering End Piece
SPENG
OO
N
DO NOT SCALE DRAWING
i
4
ItIo
4
1
MIT Mechatronics Research Laboratory
DATE
SIZE IDVASI NO.
'""'"
tv
SCALE
j
2 ICAD
2
CS102
FILE
I
1
Ibmt:I
i
Lk
A
7
8
I
I
1
6
TE INFORMATION CONTAINED IN THIS DRAWING IS THE SOLE PROPERY OF
MTand A0MRtmoCec ANY REPRODUCTION IN PART OR WHOLE WITHOUT
lEE WRITTEN PERMISSON IS PROIIT.
5
+
8
3
4
:
1
2
I
L
0
D
D
. 9
2x 4-40 TAP T.19\
C
2x 8-3 2 TAP THRU
1
C
.19K
CEs
W
wI
2x 0.13 THRU
.38
1 .19
0
B
1
I
I.
L
I
4
B
.38
.50
6.00
ITE-
PI
NO
0TFNG
ORMA-
OR DESCR
NO
PARIS
UL.ESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTIONS
+
A
DECIMALS
XX+ -(
.)DO(+.)=o
ANGLES
MIT Mechotronics Research Laboratory
DATE
+1
MATERIAL
Aluminum 5052
w
TEUN
UST
CAD GENERATED DRAWING.
DO NOT MANUALLY UPDATE
APPROVALS
EECIFIATOR
Eric Hoorau Feb 2001
Abe Schnek er
xw ENG
A
Cutting Tip Changing Station:
Shaft Holder
RNtSH
NEXTASSY
USED
ON
DO NOT SCALE DRAWING
APPCAlON
15
1
II
//
I
I
6I
6
1
APPL4IICATI2OIN
b
1
4
XlALENG
3
TI
SIE
A
NO
SCALE
ICAD FILE:
2
CS103
ISHEET
1
1
OF
I
,
8
77
6
MT
I,
4
55
THE INFORMAION CONTAINED IN THIS DRAWING IS THE SOLE PROPERTY OF
cnd A*** Phamaceutico ANY REPRODUCITON IN PART
WHOLE WITHOUT
THE WRITrEN PERMISSCTN IS PROIITED.
33
I
2
1
I
OR
D
eb
1
T1
D
C
C
~JI
-
-4
.4
5
~Q
0
-I
1.14
-4
.92
.92
.92
.92 -
.50
2x 0.13 THRU
.19
US
4..
.32
5:
5.
5x 4-40 TAP THRU
B
.50
S
6.00
PART OR
MTIM1NG NO.
ITEM
NO.
This part is made out of white DELRIN
UNLESS OTHERWISE SPECIFIED
DIMENSIONS ARE IN INCHES
TOLERANCES ARE:
FRACTONS DECIMALS ANGLES
+
.0(+ )0( +1I
A
CAD GENERATED DRAWING.
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Cutting Tip Changing Station:
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SPECIFICATION
DATE
Eric Hoorau Feb 2001
Abe Schneid er
MATERIAL
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MIT Mechatronics Research Laboratory
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Eric Hoarau Feb 2001
Abe Schnec er
Cutting lip Changing Station:
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Appendix G
The Temperature Controlled Storage Station
179
G1 Derivations for the Temperature Controlled Station
Enclosure
interior
T8
Ts
/
Q2
Insulation
HIt
Qc
Qi
Thermoelectric
cooler
Tcold
Li
Thot
Heat sink
Qh
Figure Gi: Diagram of the heat transfer in the storage system
G1.1
Heat Load Calculations
The heat gained by convection from the top is
Q2 = hA2 (T.
(G.1)
T-T)
where
h
A2
T.
T,
is the heat transfer coefficient [watts/m 2 -oC]
is the exposed top surface area [m 2
is the ambient temperature [*C]
is the temperature inside the enclosure [*C]
The heat gained through the sidewalls of the insulated enclosure is
Q, =
A, (T. - T
K
(G.2)
h
where
A]
x
K
is the external surface of the insulated enclosure [in 2
is the thickness of the insulation [m]
is the thermal conductivity of the insulation [watts/m-*C]
180
The heat released to the hot side of the thermoelectric cooler is
Qh = Qc +(Vinin)
(G.3)
where
Qe
Vin
Iin
G1.2
is the heat absorbed from the cold side of the thermoelectric [watts]
is the input voltage of the thermoelectric [volt]
is the input current of the thermoelectric [amp]
Calculation of Additional Parameters
The heat sink performance is measured in terms of its thermal resistance:
O, =
,
(G.4)
Qh
where
Thot
is the heat sink temperature [*C]
The time, t, needed to change the temperature of an object can be estimated to be
t=
P",(G.5)
where
m
C,
AT
Qave
is the weight of the material [grams]
is the specific heat of the material [cal-gram-*C]
is the temperature change of the material [*C]
is the average rate removal [cal/second]
The average rate removal is
Qe = QCDTmi
-
2
(G.6)
QCDTm
where
DT
is the temperature difference across the module [*C]
QCDTmin is the amount of heat the thermoelectric module is pumping at the initial object
temperature. DT is zero at this time and the heat-pumping rate is maximum [cal/second].
QcDTmax is the amount of heat the thermoelectric module is pumping at the final object
temperature. DT is high and the heat-pumping rate is minimum [cal/second].
181
G1.3
System Parameters
Experimental variable
Value
LI
17 [cm]
Wi
10 [cm]
H1
5 [cm]
x
2 [cm]
Ts
4 [*C]
T.
22 [*C]
Thot
42
Tcold
2 [*C]
Vin
12 [volt]
Iin
5 [amp]
K (polyurethane)
0.035 [watts/m-"C]
h (still air)
23 to 28 [watts/m 2 -*C]
h (turbulent air)
85 to 113 [watts/m 2 -oC]
Calculated Variable
Value
Al
27x10-3
[M 2 ]
A2
17x 10-3
[M2]
Q1
0.715 [watts]
Q2
7.65 [watts]
Qc at DTmin (from chart)
65 [watts]
Qc at DTax (from chart)
25 [watts]
Qave
45 [watts], 10.75 [cal/sec]
182
G2 Thermoelectric Cooler Data Sheet
U
TECHNICAL DATA SHEET
ferrotec
INTESNATIONAL TfERM0ELCTRIC INC.
THERMOELECTRIC COOLER: 127-Couples, 9.5-Amperes
T
w
Past Number
6300/127/085*
Red
I
(150 0 C Maximum Temperature)
Black
Add Suffix for Height Tolerance
*
Am *0.30mm (.010")
B= * 0.03mm (.001")
39.7mm (1.56")
Li
W =39.7mm (1.56")
(0-155")
-9mm
3
H =
135mm (5.25")
Lw=
DESCRIPThON
and/or heating applications.
This 127-couple, 8.5-amp module is a gencral purpose single-stage thermoclkctric cooler suitable for various cooling
is required. Typical
capacity
pumping
heat
high
and
used
be
is
to
source
power
This device is particularly useful where a 12-volt DC
and laboratory and scientific
application areas include consumer products; biomedical instruments; industrial, military, and electrical equipmeat
surfaces.
apparatus. The module featurcs alumina ceramic f&ce plates that clectrically isolate internal circut elements from the external mounting
Th -250C
GENERAL SPECIFICATIOnS
Tcmprature Differential (DT) at Zero Heat Load
Heal Pumping Capacity (Qc) at Zcro TemperaWurm Differmtial
Th= 50"C
Th - 900C
68
72
76
Degrees C
87
Watts
8.5
Amperes
19,2
VoltsDC
65
Maximum or Optimum Current (lopt)
Nominal Input Voltage (Vin) at optand DT - 300C
/
Th= 35'C
72
8.5
76
8.5
80
8.5
14.9
15.7
16.8
Coefficient of Performnce
Oc vs. I
"5A
50C
Th
2.0
1
--
.-
---OTO 10
D20
07.
--
30
/
Vin
Th = 50C
I
vs.
OT
127 Cpl / &SA
C=05
In -
S
1.0
to
3.
L-
50
MPER15
1
go
-
)
183
/ 85A
ss
-
O
-
-
Vin~ vs. Th
127 Cp
= 30C
C
C
G3 Assembly of the Thermoelectric Cooler Storage Station
Figure G2: The temperature controlled storage station
The sample storage station is shown in Figure G2. The steps for the construction of the tank are
shown in Figure G3. The first step is shown in Figure G3a. Four 3/8"-16, 4" hex head cap
screws are attached to the stainless steel pan with epoxy, and a thermistor is attached to the side
of the pan with a thermally conductive epoxy. The second step consists of insulating the pan
with polyurethane foam and is shown in Figure G3b. Tape was placed at the location where the
thermolectric will contact the pan. Then, the foam was applied. Once the foam had cured, it was
trimmed to the shape shown in the figure.
a)
b)
Figure G3: Construction of storage tank: a) frame only, b) with the insulation
184
a)
i
H
b)
Figure G4: Assembly of the thermoelectric base: a) top view, b) bottom view
The base of the station is shown in Figure G4. The hot side of the modules was attached to the
heat sink/fan assembly with a thermally conductive epoxy as seen in Figure G4a. The heat
sink/fan assembly was then bolted to the aluminum base plate. The temperature controller and
its knob were also bolted to the base plate. The modules are electrically in series and thermally
in parallel. The cold side of the module is coated with thermally conductive grease. The four
feet pass through the base plate and are connected to the bolts on the tank with nuts. The base
plate is pushed onto the bottom of the tank by four nuts on the feet of the assembly.
185
Appendix H
The System Software Interface
186
H1 The Interface Screens
Figure Hi: Sample of the interface control for the cutting device, transportation system, and light source
.:
Nampie o1 the interlace control for the XY stage
187