Microscopy Lab Agenda

The online class will not actually perform this lab, but you will be required to
answer questions based on the material contained in these notes and the other
microscope material present in this module.

1. Review the care and use of the microscope.

2. Identify the parts of the microscope. (See labeled microscope image and compare
to list below).
3. Explain image projection and total magnification.
1. The image light that is passing through the specimen is refracted by 2
lenses. The first lens (closes to the slide) is the objective lens. The 4
objective lenses on our microscopes have magnifications of 4x, 10x, 40x,
and 100x.
2. The second lens is the ocular lens (closest to your eye), which has a
magnification of 10x.
3. The light from the specimen is by the lenses resulting in an image being
projected to your eye looks upside-down (inverted) and backwards
(reversed).
4. The total magnification is the product of the magnification of the ocular
lens (always 10x) and the magnification of the objective lens you are using
(4,10,40, or 100).
1. Therefore the possible total magnifications with which we can
view a specimen is 40x, 100x, 400x, and 1000x.
4. View the letter “e” under the microscope with 3 objective lenses.
1. Notice the orientation of the letter ‘e’. It will appear inverter and reversed
from which you placed it under the lens.
2. Also notice that as you increase magnification, you can see less of the
entire letter, but what you do see is larger.
5. Explain focal point and depth of field. View three colored threads laying across
each other at different depths.
1. The stage moves up and down on the microscope to bring the specimen
closer and further from the objective lens. This is how we focus. There is
a specific distance from the lens that will bring the image into focus. This
is called the focal point. This is what you’re searching for when you move
the stage up & down. When moving the stage upward, structures closer to
the lens come into focue prior to structures further from the lens. The
opposite is true when you are moving the stage downward.
2. Sometimes when you view a tissue sample you are looking at several
layers of cells. Some cells come into focus while others are blurry. This
is because those cells are at different depths. Some cells look like they
have multiple nuclei. This is usually because you’re viewing several cells
on top of one another.
6. Utilize grid sheets to measure the field of view.
a. The field of view the amount of geographical space you are looking at
through the microscope. The lower your magnification, the greater the
 field of view. For example, if you’re looking at a map of Pennsylvania on
Mapquest, you may be able to see the entire state, but no specific town in
any detail. If you increase magnification of the map, you’ll see more
detail of a town, but you’ll see less of the state at one time.
b. We can calculate the field of view for all objective lenses using grid paper
with 1mm boxes:
1. We’ll view the gird paper under the 4x objective lens and count the
number of boxes in a line that passes through the center of the
circular field of view (the diameter).
1. That number of boxes is the diameter in millimeters.
2. We can then calculate the field of view of the other objective
lenses with the following formula:
1. Diameter B = (diameter A x magnification of
A)/Magnification of B
7. Explain oil immersion using the 100x objective lens.
1. The 100x objective lens provides a lot of magnification. The resolution of
this lens may not be great. A drop of immersion oil on the slide when
changing from 40x to 100x will help resolution.
8. Prepare a wet mount slide of your own cheek cells and view them under the
microscope with all 4 objective lenses.
1. We will scrape some cells from the inside of our cheeks and add them to a
drop of methylene blue stain on a slide.
2. We’ll then view the cells under the microscope and estimate the size of the
cheek cells using field of view.

Identify the following parts of a microscope (the numbers below correspond to the
labeled image(s) on the site:
1. Head
2. Ocular lens
3. Arm
4. Rotating nosepiece
5. Objective lenses (4x, 10x, 40x, 100x)
6. Stage
7. Mechanical stage
8. Mechanical stage controls
9. Slide retainer
10. Course adjustment knob
11. Fine adjustment knob
12. Condenser
13. Iris diaphragm lever
14. Light source
15. Base
16. Power switch
17. Rheostat (light intensity adjustment)