1
TABLE OF CONTENT
CONTENTS ............................................................................................................................................ 2
1. INTRODUCTION .................................................................................................................................. 4
2. MICROSCOPE HARDWARE .............................................................................................................. 5
2.1 Installation of the thermoregulated slide holder module ....................................................... 10
2.2 Condenser user guide ............................................................................................................................. 12
3. FLUORCAM7 SOFTWARE .............................................................................................................. 13
3.1 Live FluorCam7 window ....................................................................................................................... 14
3.2 General Icons, description of Live FluorCam7 window ........................................................... 15
3.3 Light source settings in Live FluorCam7 window ...................................................................... 16
3.4 FluorCam7 main menu .......................................................................................................................... 17
3.5 Description of the panel for setting the intensity of the measuring flashes, the actinic
light and super pulse ..................................................................................................................................... 18
3.6 Description of the table holder coordinates adjustment and other feature settings .. 19
3.7 Additional features in the Live window ........................................................................................ 20
4. MICROSCOPE TOUCHSCREEN SOFTWARE.............................................................................. 21
4.1 Main menu - Home .................................................................................................................................. 21
4.2 Main menu - Display ............................................................................................................................... 22
4.3 Microscope menu ..................................................................................................................................... 23
4.3.1 Turret .................................................................................................................................................. 23
4.3.2 Light..................................................................................................................................................... 24
4.3.3 Automatic.......................................................................................................................................... 25
4.2.4 XYZ........................................................................................................................................................ 26
4.4 Config menu................................................................................................................................................ 27
4.4.1 Components...................................................................................................................................... 27
4.4.2 Extras .................................................................................................................................................. 27
4.4.3 Info ....................................................................................................................................................... 33
4.5 User menu ................................................................................................................................................... 33
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4.5.1 User selection .................................................................................................................................. 33
4.5.2 User config ........................................................................................................................................ 34
4.5.3 Admin .................................................................................................................................................. 37
5. SPECTRUM SOFTWARE ................................................................................................................. 38
5.1 Spectrum and FluorCam7 software .................................................................................................. 40
5.2 Spectrum – Spectrometer software Main menu.......................................................................... 41
5.3 Spectrum – Preview window .............................................................................................................. 42
5.4 Spectrum – Settings for the Run mode measurement .............................................................. 43
5.5 Spectrum – Run mode measuring ..................................................................................................... 44
5.6 Spectrum – Get Spectrum mode measuring.................................................................................. 45
5.7 FluorCam7 – Protocol set-up for parallel spectrometer measurements .......................... 46
5.8 FluorCam7 – Protocol modification ................................................................................................. 47
6. REFERENCES ..................................................................................................................................... 48
3
1. INTRODUCTION
Dear customers,
This manual is designed to provide additional information for the specific use of
the multi-color MicroFC. For general operation of the FluorCam system please refer to
FlurCam Instruction manual.
Please note that additional instructions for the operation of the MicroFC
integrated Spectrometer is included.
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2. MICROSCOPE HARDWARE
FIGURE 2.1: The entire assembly.
(1) Microscope, (2)Microscope Touchscreen, (3)FluorCam, (4) Light Sources, (5) Power source for
FluorCam and Light sources, (6) Spectrometer, (7) Microscope power source.
FIGURE 2.2: Microscope.
(1) Eyepiece, (2) Reflector turret (two objectives: 20x, 40x), (3) Transmitted bottom light, (4)
Touchscreen (see chapter 4), (5) FluorCam, (6) Control of the stage to the sides, (7) Focus, control
of the stage to up/down.
5
FIGURE 2.3:Microscope touchscreen (see chapter 4).
FIGURE 2.4: FluorCam.
(1) FluorCam device, (2) USB cable connected to the computer, (3) Power source cable (see fig. 2.6).
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FIGURE 2.5: Power source of the FluorCam and of the Light sources.
(1) Power switch, (2) EPI – illumination: Indicator of flash lights, actinic light, super pulse and
STF, (3)Transillumination: Indicator of transmitted light.
FIGURE 2.6: Power source of the FluorCam and of the Light sources backside view.
(1) Power source cabel, (2) Flash light source cabels, (3) Actinic, super pulse and STF light source
cables, (4) Transmitted light source cables, (5) FluorCam cable, (6) Spectrometer cable,
(7) Microscope cable.
7
FIGURE 2.7: Actinic, super pulse, STF light sources.
Parallel positioned relative to the microscope.
FIGURE 2.8: Flash light sources.
Vertically positioned relative to the microscope.
8
FIGURE 2.9: Transmitted light sources.
Positioned on the bottom of the microscope.
FIGURE 2.10: Backside view of the microscope.
(1) Cable connected with the power source of the FluorCam and of the Light sources.
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FIGURE 2.11: Spectrometer device.
(1) LED indicators of the running the device (Power), the cooling of the device (Cooling)and the
hardware errors (HW Error). The power switch is located on the backside of the device. (2) Fiber
optic cable.
2.1 Installation of the thermoregulated slide holder module
FIGURE 2.12: Installation of the thermoregulated slide holder module.
Remove by screwing the bolts the classic slide holder before the instalation. Prepare the slideholder
on the stage, under the microscope should be gap.
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FIGURE 2.13: Installation of the thermoregulated slide holder module 2.
Place the thermoregulated slide holder modul between two bolts.
FIGURE 2.14: Installation of the thermoregulated slide holder module3.
Tighten both bolts.
For the further information about the thermoregulated slide holder please see the
Water cooling circuit installation manual.
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2.2 Condenser user guide
FIGURE 2.15 Condenser.
For the objective with magnification 10x, adjust the transmitted light/bright-field
according to Koehler.
Use front optics in place.
After adjusting bring front optics on condenser out of place by lever on the picture for
homogeneous lighting of the specimen.
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3. FLUORCAM7 SOFTWARE
Before measuring:
1. Turn on the microscope power source.
2. Turn on the microscope (the button is located on the left side of the
microscope stand).
3. Turn on the FluorCam power source (FluorCam microscopy version control
unit for lights).
4. Switch ON the PC.
5. Launch the FluorCam7 software.
The microscope starts calibrate its stage after launching the FC7 software. The
stage should be set in the center position (under the objective is a gap), otherwise, the
blocking can occur if thermoregulated slide holder module is used.
WARNING! If you use thermoregulated slide holder module do not place the
microscope slide under the objective before the calibration of the stand zero position is
finished. If objectives are positioned above the slide, the slide will break down!!!
Please note that it is not possible to change the objective (from 20x to 40x and
back) if thermoregulated slide holder module is used and the microscope stage is too
close to the objective (in the upper position). Please follow the instructions to position
the table for imaging under different objectives if thermoregulated slide holder module
is used:
1. Lower the stage.
2. Change the objective.
3. Set the position of the stage as required.
WARNING! For obtaining the optimal signal, please, do not change the binning
2x2.
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3.1 Live FluorCam7 window
FIGURE 3.1: Live FluorCam7 window.
The life window can be divided into 6 parts. Image of the sample (1), Light sources panel (2) for
switch on and off all available light for the given instrument version, Global Light Settings panel
(3) where is possible to change the light intensity, Camera panel (4) for adjusting camera settings,
Flashes and Actinic/Super panel (5) for changing light intensity and Coordinate, diaphragms,
Reflector turret and Side port settings (6).
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3.2 General icons, description of Live FluorCam7 window
FIGURE 3.2: General icons, description of Live FluorCam7 window.
Clicking the magnifying and de-magnifying icons (1), one can increase and decrease the size of the
image window on the computer screen. The mouse wheel can be used as well. The current zoom is
shown in the field Zoom (2). By clicking the numeric scale field (3), the scaling bar is supplemented
by a numeric scale bar. By clicking anywhere on the color / gray scale bar (4), one can change the
color scheme of the image (5). Device (6) displays the present FluorCam device and its Resolution
(7). The FluorCam ID and Version are automatically recognized and displayed in Version (8). (9)
shows the type of a present camera.
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3.3 Light source settings in Live FluorCam7 window
FIGURE 3.3: Light source settings in Live FluorCam7 window.
Top right Light Sources panel (1) is used to switch on and off all available light for the given
instrument version. In the given example the measuring flashes, the actinic light (Act1), the super
pulse (Super), the far-red light (FAR) and the transmitted bottom light (Trans) is used. In the
Global Light Settings panel (2) it is possible to change the light intensity by pulling the trigger (3)
or by writing the exact value in the field under the given light trigger (4). Important: It is
recommended to set the value for the lights available to 100% and change the intensity in the panel
Flashes and panel Actinic/Super .The shutter El.Shutter(x) (5) and Sensitivity (6) can be adjusted
manually by pulling the trigger. The actual adjusted setting of the live window can be transmitted
to the protocol settings by confirming it with USE button (9). The average or none software filter
can be chosen in Filter (7) and the contrast mode can be checked in Contrast (8).
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3.4 FluorCam7 main menu
FIGURE 3.4: FluorCam7 main menu.
‚ Experiment‘ menu (1) serves for saving and re-opening measured files and for exporting
acquired results. ‚Protocols‘ menu (2) is for opening, modifications and saving of measuring
protocols. ‚Setup‘ (3) enables advanced settings, calibrations and updating the device. ‚Help‘ (4)
gives information about the software and producer. Click the 'Wizard icon‘ (5) to open pre-defined
protocol menu and wizard protocol menu available for the given device. (6) Run trigger initiates
execution of the experiment (starting from compilation of the protocol, if not done in the protocol
window). (7) 'Stop‘ interrupts execution of the experiment. After the Stop is used, the experiment
can be re-started (but not resumed). (8) Multiple run trigger is available in spoftware upgraded
versions and is used for advanced protocol settings and automated multiple protocol
measurements.
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3.5 Description of the panel for setting the intensity of the measuring
flashes, the actinic light and the super pulse
FIGURE 3.5: Description of the panel for setting the
intensity of the measuring flashes, the actinic light
and the super pulse.
In the panel Flashes (1) it is possible to change the
measuring flash intensity, in the table Actinic/Super
(2) the intensity of the actinic light and the super pulse
can be changed.
For switching on and off the chosen light (4), the icon
below the given light (3) must be checked.
The intensity of light can be changed by pulling the
scale trigger (5) or by writing the exact value in the
field below the given light (6).
.
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3.6 Description of the table holder coordinates adjustment and other
feature settings
FIGURE 3.6: Description of the table holder
coordinates adjustment and other feature settings.
In the panel Coordinate the position of the holder stage
can be changed. The trigger (1) is coarse focus of the
stage for approximate positioning of the stage. The circle
trigger (2) is used for fine positioning of the stage and for
exact plane focusing of the imaged object. Use the mouse
to turn with the black dot on the wheel to the left or right
for the fine focusing to the correct object plane . Current
position is shown in numbers below the wheel.
In the window (3) it is possible to save, load and delete the
exact position of the stage holder in which given object is
in focus .
The diaphragm of reflector field can be adjust by the
trigger RefFieldDiaph(4) and the diaphragm of
transmitted field by the trigger TransFieldDiaph (6).
The intensity of the halogen lamp is set by the trigger
LampHALIntens (5).
The different filters available in the filter wheel can be
chosen by the scrolling window Reflector turret (7). The
numbers of the filters correspond with the labels on the
front of the microscope.
The shutter of the reflector can be turn on and off by Refl.
Shutter (8).
Is also possible to connect other devices with the
microscope (e.g. spectrometer). In this case the Side
port (9) should be turn on.
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3.7 Additional features in the Live window
FIGURE 3.7: Additional features in the Live window.
The image shown in the image window can be captured as a snapshot by „camera icon” (1). „Hot“
pixels can be removed by „ ambulance icon”(2). Scrolling menu (3) shows the type of a camera.
SnapShot(4) is a snapshot window, which appears after clicking the „camera icon” (1). „Camera
icon” (1) enables taking an image snapshot and saving the image (Save in window (4)) as a bitmap
or exporting it as *.fcimg (Export in window (4)).
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4. MICROSCOPE TOUCHSCREEN SOFTWARE
The microscope touchscreen turn on automatically when the microscope is turning
on. The button for switch on/off the microscope is situated on the left side of the
microscope stand. The display is controled by touching of the fingers or by the stylus. If
some of the buttons on the touchscreen are activated, they have a grey color, otherwise,
they are blue. In the submenu, if some panel is shown, it has a light blue color, otherwise,
the panel is grey.
4.1 Main menu – Home
FIGURE 4.1: Main menu – Home.
In the menu (1) on the left part of the display is possible to browse between different settings of the
microscope. In the middle are displayed current settings of the microscope. Which user (and his
settings) is currently using the microscope is visible in the frame User (3) (it also can be set as
“Default”).The exact position of the stage , which was used and saved previously ,can be load by
clicking at the button Load Position(4). Reflected and transmitted illuminations are possible to
turn off/on by clicking the buttons RL Illumination (5) and TL Illumination (6). For better
visibility of a sample use the button Make it Visible! (7). The information about currently used
type of the objective (8), the reflector filter (9) and the intensity of TL Illumination (10) are visible
on the bottom of the display. By clicking the button Home (11) during the setting of the microscope
parts, an user is coming back to the main menu. In folder Microscope (12) is possible to set turret,
light, automatic settings and position of the stage. In Config (14) the different components of the
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microscope can be set. In the folder User (14) the personal favorite settings of every user of the
microscope can be selected. The brightness and turning off the display can be chosen in folder
Display (15). If the buttons have blue background it means, that they are inactive. The activated
buttons have grey background.
4.2 Main menu – Display
FIGURE 4.2: Main menu – Display.
The brightness of the display is showed in Background brightness (1) and can be changed by
clicking the arrows (2). The display is possible to turn off by using the button Display off (3).
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4.3 Microscope menu
4.3.1 Turret
FIGURE 4.3.1: Microscope menu – Turret – Contrast:
In panel Contrast is possible to turn on/off the contrast manager. If Contrast Manager Reflector
(1) is set to Empty, than RL illumination is turn off and TL illumination is turn on and the contrast
mode is not used. The same happens, if the Contrast manager Transmitted Light (2) is set to BF
(Brightfield), that means, that contrast mode is switched on.
FIGURE 4.3.2: Microscope menu – Turret – Reflector:
The selected filter (Chl, DAPI, Multiband, etc.) can be set in panel Reflector. The filter position
number (1) corresponds with the number on the label, which is situated on the front of the
microscope. It is also possible activate the Contrast manager (2). The contrast, which is currently
set, is visible in the table Current Contrast (3).
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4.3.2 Light
FIGURE 4.3.3: Microscope menu – Light – Light Path:
This panel describes a light path in the microscope. The panel is divided in Top port part (1) and
Side port part (2). By clicking blue square button (3) is possible to change the light path from Tube
to Side port and back.
FIGURE 4.3.4: Microscope menu – Light – Illumination:
The TL intensity of the halogen lamp HAL TL (1) can be set by clicking the arrows (2). The current
setting is showed under the bar scale.
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FIGURE 4.3.5: Microscope menu – Light – F/A TL:
The TL diaphragm Field Stop (1) can be set manually by clicking the arrows (4) or automatically
by using the buttons Prev – 112 mm (2) and FOV – 25 mm (3).
4.3.3 Automatic
FIGURE 4.3.6: Microscope menu – Automatic – Soft Keys:
Is possible to set and use more functions by clicking one single button Function in Soft Keys panel.
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FIGURE 4.3.7: Microscope menu – Automatic – HW Settings:
Hardware setting of the microscope can be set automatically by clicking the certain button, for
example: -1-.
4.3.4 XYZ
FIGURE 9: Microscope menu – XYZ – Position:
The position of the stage in the direction Z (up/down) can be sat at table Z-Position (1). The
setting of the zero position can be set manually (actual position is 0) by Set Zero man (2) or
automatically (the stage changes its position to the end position, which is set to 0) by Set Zero auto
(3). A simple distance measuring between two positions can be provided and showed in Delta Z
frame (4). The measuring is activated by clicking the Start button. The set position can be saved by
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Save Position (5) or be deleted by Delete (6). The certain position can be set by clicking one of the
buttons (Pos. 1) in the frame Position (7).
4.4 Config menu
4.4.1 Components
FIGURE 4.4.1: Config menu – Components – Objectives:
The description of objectives is in the Objectives panel. The position is given by the blue frame (1)
and the type, magnification level and the other information are in the white frame (2).
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FIGURE 4.4.2: Config menu – Components – Reflector:
The reflector turret information can be configured in this panel. The position of the turret is in the
blue frame (1) and the information about the reflector turret in the white frame (2). To see all the
information the configuration must be done.
FIGURE 4.4.3: Config menu – Components – Focus:
The focus speed in the frame Focus speed (1) can be set for the selected objective (2) by clicking
the white frame (3). Parfocality can be activated and adjusted (configuration manager) in (4).
Stage drop (moving down) is possible to turn on/off in (5) and is also possible to set the distance of
the drop in Drop Distance (6). The stage drop can be managed by the Software limit (7).
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FIGURE 4.4.4: Config menu – Components – Camera ports:
The panel Camera-ports is divided into two parts: In part Adapter (1) it is possible to set camera
adapter ports by clicking french key image (it must be white, if is grey, it is not possible to make any
changes – please note, that only administrator can do the changes) (3). The information about
current settings is showed in the white frame (4). In the part Splitter (2) is possible to set
beamsplitters in the same way like in the part Adapter.
FIGURE 4.4.5: Config menu – Components – Misc:
In this panel the type of TL or RL illumination, turret mot., FOV eyepiece, autofocus, manual
condenser, settings of tube light turret and reflected light plugin component can be configured by
clicking the icon of french key (please note, that only administrator can do the changes).
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4.4.2 Extras
FIGURE 4.4.6: Config menu – Extras – Light manager:
In this panel is possible to turn on the light manager, which automatically sets the brightness by
Light Manager Mode (1).The light manager values can be defined by user or the values can be set
as default (factory settings) by Load Light Manager Values (2).
FIGURE 4.4.7: Config menu – Extras – Contrast manager:
Turning on/off the contrast manager, which automatically sets the contrast.
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FIGURE 4.4.8: Config menu – Extras – Oil Stop:
If the Oil Stop manager is set to On, the drought objective is not dip into the immerse oil when user
is changing the immerse to the drought objective.
FIGURE 4.4.9: Config menu – Extras – Dazzle Protect:
The dazzle protection is provided by switching on the buttons Dazzle Protection (global), Dazzle
Protection Reflected Light and Dazzle Protection Transmitted Light (1).
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FIGURE 4.4.10: Config menu – Extras – Ethernet:
In this panel is possible to connect the microscope to the Ethernet. For writing use the keyboard (1).
FIGURE 4.4.11: Config menu – Extras – Misc:
In the Misc folder is possible to do the TFT calibration, if the display is not reacting exactly on a
touch.
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4.4.3 Info
FIGURE 4.4.12: Config menu – Info – Firmware:
This panel contains information about microscope firmware (serial numbers (1), date of
manufacture (2)).
4.5 User menu
4.5.1 User Selection
FIGURE 4.5.1: User menu – User Selection – User:
In panel User is possible to choose certain user and his settings clicking User 1 (2) or work with the
default settings choosing Default (1).
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4.5.2 User Config.
FIGURE 4.5.2: User menu – User config. – HW Settings:
Hardware setting of the microscope can be set by an user by clicking the certain button, for
example: -1-.
FIGURE 4.5.3: User menu – User config. – User Settings:
In this panel is possible to change the password to the user profile with his favorite settings.
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FIGURE 4.5.4: User menu – User config. – Buttons left:
In Buttons Left panel is possible to assign different function to the buttons on the button wheel (3)
(focus in Z plane and buttons on the microscope stand) by clicking the blue frame (1). The selected
option is displayed in the white frame (2). Warning! Only the administrator can make the changes.
FIGURE 4.5.5: User menu – User config. – Buttons Right:
In Buttons Right panel is possible to assign different function to the buttons on the button wheel
(1) (focus in Z plane and buttons on the microscope stand) by the same procedure like in fig. 4.5.4 .
Warning! Only the administrator can make the changes.
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FIGURE 4.5.6.: User menu – User config. – Buttons left:
In Docking Station panel is possible to assign different function to the buttons on the button wheel
(1) (docking station button and buttons on the microscope stand) by the same procedure like in fig.
4.5.4. Warning! Only the administrator can make the changes.
FIGURE 4.5.7: User menu – User config. – Favorite page:
An user can pick his favorite page in the scrolling menu Select the favorite page (1). For the
moving in the menu serve blue arrows (2). For confirm the choices use the button Apply selected
page as Favorite page (3).
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4.5.3
Admin
FIGURE 4.5.8: User menu – Admin: To the folder Admin is access only after entering the password
in the frame Password (5 characters) (1). For entering the password use the keyboard (2).
FIGURE 4.5.9: User menu – Admin – Settings:
In this panel the Administrator can change his password by clicking Change administrator
password (1) and set the language and system start-up configuration (2).
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FIGURE 4.5.10: User menu – Admin – User manager:
Administrator can set default (1) and users settings (2) in User Manager.
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5. SPECTRUM SOFTWARE
Quick instruction:
1. Turn on the spectrometer power source.
2. In the FluorCam7 software in the lower-right corner the Side port should
be ON.
3. Choose protocol in FluorCam7 software which you want to use for your
experiment.
4. Modify the protocol (see fig. 7 and 8).
5. Open Spectrum software and select the measuring mode (see fig. 2).
6. Set the spectrum measuring options (only if the Run mode measurement
is selected, see fig. 4).
7. Launch the spectrum measuring (button Run).
8. Launch the experiment in FluorCam7 software.
FIGURE 5: Optical Spectometer
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5.1 Spectrum a FluorCam7 software
FIGURE 5.1: Spectrum and FluorCam7 software:
It is possible to measure fluorescence data using FluorCam7 software (1) and measure spectra
using Spectrum software (2) simultaneously. For this option is necessary to set the Side Port (3) to
ON.
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5.2 Spectrum – Spectrometer software Main menu
FIGURE 5.2: Spectrum – Spectrometer software Main menu:
After launching the Spectrum software, the main Measuring window is opened. The spectral
results can be obtained in two different modes: manual and automatic based on pre-defined
settings. For the manual mode, Get Spectrum (2) icon is used. In manual mode always one
spectrum is measured and is displayed in the Measuring window after Get Spectrum icon is used.
Please note that Preview spectrum icon must be checked (4) to view the spectrum. For the
continuous automatic measurement mode use the Run icon (1). If Run icon is checked spectra are
measured continuously according to pre-defined settings (please check fig. 5.4 for further
explanation). Please note that this mode is used only for Fluorcam protocol measurements. To
obtain spectra during Fluorcam protocol measurements, the Fluorcam protocol must be modified
before launching. It is necessary to define time for spectra acquisition as it corresponds to given
phase of kinetic fluorescence measurement (please refer to fig. 5.7 and 5.8 for further explanation).
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5.3 Spectrum – Preview window
FIGURE 5.3: Spectrum – Preview window:
If the Preview Spectrum (1) is unchecked, it won’t be possible to see the spectrum graph the
window (2). By clicking the Stop (3), the measuring of the spectrum is stopped.
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5.4 Spectrum – Settings for the Run mode measurement
FIGURE 5.4: Spectrum – Settings for the Run mode measurement:
Before the measurement it is important to set the Integration time (1) and the Device
parameters (2) in the scrolling menu Setup. For measurement it is recommended to use Control
Synchronization mode and External Trigger Pulse as a Sensor Work Mode. Average number
determines averaging of a certain number of samples {pulses). If the Run mode measuring is
selected, it is necessary to set Number of samples to be measured (3). If zero is chosen, there is no
limitation in the number of obtained spectrums. It is recommended to use zero value during
Fluorcam protocol measurements. In the example shown above, each 30 ms during Fluorcam
protocol measurement one spectrum is obtained. In total 10 spectrums are measured (please see
Count number (4)).
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5.5 Spectrum – Run mode measuring
FIGURE 5.5: Spectrum – Run mode measuring:
To start the spectra acquisition during Fluorcam protocol measurements please follow next steps:
1. Launch the experiment in FluorCam7 software, adjust the measuring protocol and define
time in the Fluorcam protocol for spectra acquisition (please refer to Fig. 5.7 and 5. 8 for
further details).
2. Prior initiating the Fluorcam protocol run it is IMPORTANT to click on the Run button in
Spectrum software. It is possible to see, how many spectrums were measured (1).
3. In the scrolling menu Data it is possible to export obtained spectrum results (2) to the
Notepad (4) or clear them and start new measuring. It is not possible to view the spectrum
graph in the Run mode measurement setting.
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5.6 Spectrum – Get Spectrum mode measuring
FIGURE 5.6: Spectrum – Get Spectrum mode measuring:
If the measuring is started by clicking Get Spectrum button (1), the spectrum graph is shown in the
window (2). The list of measured spectrums is displayed in the panel (3). The spectrum which is
checked (4) is currently visible in the measuring window (2). Spectrum can be also checked (10),
unchecked (11), reloaded (12) and deleted (13) by clicking at these buttons. It is also possible to
save them and after that load them again or export results of all the spectrums or export only the
selected ones. This can be managed in scrolling menu Data (5).
In menu (6) one can configure lights and manual scale. In the menu Math (7) it is possible to do
mathematical calculations with the spectrums (not available).
If Logarithmic (8) is checked, the scale in the graph window (2) is logarithmic. It’s also possible to
use Auto Integration function (9).
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5.7 FluorCam7 – Protocol set-up for parallel spectrometer
measurements
FIGURE 5.7: FluorCam7 – Protocol set-up for parallel spectrometer measurements:
In the header of every protocol, which will be used to obtain fluorescence and spectrofotometric
data, there must be definition for the use of spectrometer defined as: include spectrometer.inc (1).
This definition is automatically pre-defined in all PSI designed protocols and wizards for the
MicroFC systems with integrated Spectrometer.
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5.8 FluorCam7 – Protocol modification
FIGURE 5.8: FluorCam7 – Protocol modification:
To obtain fluorescence and spectrofotometric data it is essential to define phase of the fluorescence
kinetic measurement in which the spectrum should be automatically in pre-defined intervals
recorded. The user must define in which phase of the fluorecence transients of the given Fluorcam
protocol measurement the measurement of the spectrum begins. In this example the initiation of
spectrum with spectrometer is set at the beggining of Kautsky effect measurement. In the header of
the Kautsky effect should be formula (1) which describes the beggining of the measurement c1 (2),
the step c1+1 second (3), the end ALPeriod (4) (in this case, the measuring of the spectrum will last
all Kautsky effect measuring) and the command spec_sample (5) (which triggers the spectrometer
measurement). It is possible to define multiple time points in the Fluorcam protocol when
spectrometer measurements will be initiated.
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6. References
References: Carl Zeiss spol. s.r.o.: Prirucka pro obsluhu Axio Imager primy mikroskop.
20/12/2005, translation 21/01/2008. B 46-0078 cz. p 87-110.
Manual Version: 2013/12
© PSI (Photon Systems Instruments), spol. s r.o.
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