1. Microscopy lecture: Text Chapter 3 (supplement pg 39)
Lab 2 Goals and Objectives:
Microscopy lecture: Text Chapter 3 (supplement pg 39)
Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in supplement pg 41)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion
Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (supplement pg 42)
Microscope Worksheet (supplement pg 43)
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 supplement pg 45-46

2. Microscopy Text book Chapter 3
Lab Notes
Supplemental Packet page 39

4. Light Microscope

6. Brightfield Darkfield Phase contrast D.I.C.

7. Fluorescence Microscopy

8. D.I.C.
Confocal

9. Electron Microscopy

10. Scanned-Probe Microscopy
DNA
Bacterial Surface Protein

11. Positive Staining
Negative Staining

12. Gram Stain
Acid Fast Stain

13. Capsule Stain
Special Stains

14. BIO244 Microscopy
Students will need:
microscope and cord
(sign out number, use all semester)
1 “e” slide
1 stage micrometer
immersion oil: share one bottle per student pair
lens tissue & methanol: share per student pair
lab manual or photocopied pages (Exercises 1 & 5)
laboratory supplemental packet:
Guide to Using the Microscope (page 41)
BIO244 Microscope Worksheet (page 43)
Measurements using the Ocular Micrometer (page 42)

15. Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in supplement pg 41)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion

16. Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (supplement page 42)

17. Make sure left ocular is turned fully counter-clockwise
Start with 10X objective (find it with 4X objective first!)
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

18. 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?

19. 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

20. 4. Repeat for the 40X objective:

21. 0.1mm = 40 “units”
1 unit = 0.1mm ÷ 40
1 unit = mm
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

22. 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

23. Dinoflagellate 4Xobjective (40X) 40Xobjective 10Xobjective (400X)
Czura 2005

24. Lilium Pollen 4Xobjective (40X) 40Xobjective 10Xobjective (400X)
Czura 2005

25. Frog Blood 4Xobjective (40X) 40Xobjective 10Xobjective (400X) (100X)
Czura 2005

26. Streptococcus/Enterococcus faecalis
40X objective
(400X)
100X objective
(1000X)
Czura 2005

27. Lilium Pollen
Dinoflagellate
Coccus Bacteria
Frog Blood

28. Microscopy lecture: Text Chapter 3 (supplement pg 39)
Lab 2 Goals and Objectives:
Microscopy lecture: Text Chapter 3 (supplement pg 39)
Exercise 1: Brightfield Microscopy
Learn how to use the microscope (guide in supplement pg 41)
Diopter and interocular adjustments
Light adjustments (iris diaphragm)
Oil immersion
Exercise 5: Microscopic Measurements
Calibrate the ocular micrometer (supplement pg 42)
Microscope Worksheet (supplement pg 43)
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 supplement pg 45-46