BIO244 Lab 1 Microscopy
Students will need:
• microscope and cord (sign out number, use all semester)
• 2 sterile agar plates, stack and tape to keep lids shut
• 4 sterile swabs
• 1 vial sterile water
• 1 “e” slide
• 1 stage micrometer
• immersion oil: share one bottle per student pair
• lens tissue & methanol: share per student pair
• laboratory supplemental packet
• lab manual or photocopied pages
• BIO244 Microscope Worksheet
• Chapter 3 Notes outline
Microscopy
Text book Chapter 3
Lab Notes
Light Microscope
Brightfield
Phase contrast
Darkfield
http://www.microscopy-uk.org.uk/index.html?http://www.microscopy-uk.org.uk/intro/illu/illu.html
D.I.C.
Fluorescence Microscopy
D.I.C.
Confocal
Electron Microscopy
Scanned-Probe Microscopy
DNA
Bacterial Surface Protein
Positive Staining
Negative Staining
Gram Stain
Acid Fast Stain
Capsule Stain
Special Stains
Lab 1 Goals and Objectives:
Microscopy lecture: Text Chapter 3
Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in suppl. packet p.51)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion
Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (suppl. packet p.52)
Microscope Worksheet
Measure the provided specimens
Complete the microscope worksheet and turn it in
Microscope Measurement Concept Practice Homework: complete
measurement practice sheet in supplemental packet, self check:
answers in packet (p. 53-54)
Exercise 7: Ubiquity of Bacteria (Replacement activity in packet p. 55)
Homework: “Testing Household Surface” handout,
Take home plates, swabs, and sterile water
READ DIRECTIONS CAREFULLY! then do experiment
Exercise 7: Ubiquity of Bacteria (Replacement activity in packet p. 55)
Homework: “Testing Household Surface” handout,
Take home plates, swabs, and sterile water
READ DIRECTIONS CAREFULLY! then do experiment
Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in suppl. packet p.51)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion
Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (suppl. packet p.52)
1. Make sure left ocular is turned fully counter-clockwise
2. Start with 10X objective
3. Get stage micrometer (2mm ruler on slide) in focus: you will see the
ocular micrometer with labels in multiples of 10 (0, 10, 20, 30) and
the stage micrometer with labels in multiples of 0.1 (0.0, 0.1, 0.2)
stage
micrometer
ocular
micrometer
3. Line up the zero lines of the two micrometers (you can turn the top of
the ocular to position the ocular micrometer and the mechanical stage
controls to move the stage micrometer)
At 100X total magnification they should line up as shown below
“0.1” on stage micrometer is 0.1mm on that actual 2mm ruler. This
distance corresponds to 10 “units” (spaces between lines) on the ocular
micrometer (0-10): the 0.1 line of the stage ruler lines up with the 10 line
on the ocular one. How long in millimeters is one unit on the ocular
micrometer with the 10X objective in place?
0.1mm = 10 “units”
1 unit = 0.1mm ÷ 10
1 unit = 0.01mm
to convert mm to µm, multiply by 1000
When the microscope is set to 100X total
magnification, one unit on the ocular
micrometer is worth ________µm
4. Repeat for the 40X objective:
0.1mm = 40 “units”
1 unit = 0.1mm ÷ 40
1 unit = 0.0025mm
to convert mm to µm, multiply by 1000
When the microscope is set to 400X total
magnification, one unit on the ocular
micrometer is worth ________µm
When the microscope is set to 100X total
magnification, one unit on the ocular
micrometer is worth ____10____µm
When the microscope is set to 400X total
magnification, one unit on the ocular
micrometer is worth ____2.5____µm
When the microscope is set to 1000X total
magnification, one unit on the ocular
micrometer is worth ____1____µm
Dinoflagellate
10Xobjective
(100X)
Czura 2005
4Xobjective
(40X)
40Xobjective
(400X)
Lilium Pollen
10Xobjective
(100X)
Czura 2005
4Xobjective
(40X)
40Xobjective
(400X)
Frog Blood
10Xobjective
(100X)
Czura 2005
4Xobjective
(40X)
40Xobjective
(400X)
Streptococcus/Enterococcus faecalis
40X objective
(400X)
Czura 2005
100X objective
(1000X)
Dinoflagellate
Lilium Pollen
Frog Blood
Streptococcus faecalis
Lab 1 Goals and Objectives:
Microscopy lecture: Text Chapter 3
Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in suppl. packet p.51)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion
Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (suppl. packet p.52)
Microscope Worksheet
Measure the provided specimens
Complete the microscope worksheet and turn it in
Microscope Measurement Concept Practice Homework: complete
measurement practice sheet in supplemental packet, self check:
answers in packet (p. 53-54)
Exercise 7: Ubiquity of Bacteria (Replacement activity in packet p. 55)
Homework: “Testing Household Surface” handout,
Take home plates, swabs, and sterile water
READ DIRECTIONS CAREFULLY! then do experiment