1.

Fundamentals of Microscopy

Kyle Dunn

9/28/2021
Instructions:
1. Please read pages 1-9 of the investigative manual for the “Fundamentals of Microscope
lab”.
a. Once you have done so answer the prelab question BEFORE completing any of
the lab’s activities.
2. Once you have completed the prelab questions proceed to the activities of the lab. For
the microscope lab the instructions for the activities are found in this document (below)
and also the “User Manual” that comes with your lab kits microscope and has been
attached to the assignment in D2L.
a. As you read through the instructions for completing each activity make sure you
also:
a.i. Complete any instructions (append photos, etc) below.
a.ii. Answer all the questions below.
3. Here is a video that will introduce you to the lab and its main concepts. The student is
encouraged to watch it.
a. Fundamentals of Microscopy

Prelab Questions
1. State three differences in the use and design of a stereomicroscope and compound light
microscope.

Stereomicroscopes are generally used to view larger objects, such as small insects for
example, while compound light microscopes are for much smaller objects which may not
be visible to the naked eye. Because of this, stereomicroscopes and compound light
microscopes look different as well. Compound microscopes have an area for a slide that
can be moved, while stereo microscopes typically have much more space, allowing for
larger objects to be placed. Stereo microscopes often also have two eyepieces to look
through, while compound ones have a single eyepiece. Additionally, the way that lenses
are arranged to refract light varies between these two as well.

2. Fill out the following table. Note you will not look at these through a microscope but
rather are making a prediction based on the magnification stated in the left-hand column
of the table. For “Depth of Field” you can state “more shallow than fly leg” or “less
shallow than fly leg” for the top row. For the bottom row state “more shallow than fly
wing” or “less shallow than fly wing”. Finally, for the “Image appears ‘darker’ or
‘lighter’” State this in relationship to each other. So, for the top row you could say
“darker than fly leg” or “lighter than fly leg” and for the bottom row you could say
“darker than fly wing” or “lighter than fly wing”?

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 Image appears
Total Magnification Depth of Field
“darker” or “lighter”
Fly wing – 6x ocular 120x Less shallow than fly Lighter than fly leg
lens & 20x objective leg
lens
Fly leg – 6x ocular 240x More shallow than Darker than fly wing
lens & 40x objective fly wing
lens

3. For the scenarios presented in the table above, how can more light be directed through the
object to make for a better image?

If using a compound microscope, there are several options available. Assuming the
bottom illuminator of the microscope is already in use, you can also activate the overhead
illuminator to shine additional light on the sliders. Finally, you can adjust the disc
diaphragm to modify the amount of light being transmitted from the bottom illuminator.

Activity 1
1. To complete activity 1 you will need to obtain the user manual that came with the
microscope in your lab kit (and is attached to the assignment in D2L). Read pages 1-6.
Proceed to step 2.

2. Read and complete the instructions on pages 7-9 of the microscope’s user manual. Once
you have done so you are now ready to complete activity 1 starting at step 3 below.

3. In this activity we are going to be observing how changing the magnification of

observation will impact the amount of light coming into the lens. Given this read all of
steps a.- g. below and then record a purpose and hypothesis for the experiment. DO NOT
perform any of the experiment prior to recording the purpose and hypothesis.

a. Select one of the three prepared slides from your microscope’s starter kit.

b. Place this slide on the stage and clip it into place.

c. Rotate the disc diaphragm to the setting that allows the least amount of light to
reach the stage.

d. Select the 4x objective and focus your microscope on the slide currently on the
stage. Take an image of the slide using your phone and insert it in your report
below. Here is a video to help you take an image with your phone through your
microscope: Take a microscope image with a phone

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 e. Now select the 10x objective and repeat step “d.” from above. DO NOT change
the setting on the disc diaphragm.

f. Now select the 40x objective and repeat step “d.” from above. DO NOT change
the setting on the disc diaphragm.

g. Finally, go back to step d., e., and f. and change the disc diaphragm to get the best
view of the slide possible. Post the images below.

Purpose statement: (This should be the question the experiment is attempting to address. It
should be written as a question.)

Will increasing the magnification of the microscope increase or decrease the amount of light
coming in through the lens?

Hypothesis statement: (This should be an “if/then” testable prediction that addresses the
question/purpose of the lab.)

If the magnification of the microscope is increased, then the amount of visible light will decrease
as magnification increases.

Evidence/Claim statement: (This should be a statement regarding whether your hypothesis was
supported or refuted and what data/evidence allows you to make this claim.)

It appears that my hypothesis was correct. As the magnification increased, the amount of visible
light decreased as well. This is evident from my own personal observations, as well as the
attached photographs where you can observe a decrease in brightness as magnification increases.

Reflection statement: (This should be a statement of what you learned, how your understanding
changed, if you have new questions, and what connections can you make between the lab and the
content in the book and other assignments.)

While increasing the magnification did in fact decrease the brightness, I found that playing with
the disc diaphragm did not make much of a difference in clarity. This may be due to the
microscope I am using, but there was little difference in quality observed. While I would say that
things were slightly more clear on the brightest setting of the disc diaphragm, it doesn't appear
this was conveyed at all through the photographs, which the two pictures looking seemingly
identical. I wonder if this the camera simply didn't pick up the differences, or if I was biased in
my observation, and expected the clarity to increase as I increased the brightness.

Photo 1 – Activity 1

Insert the image(s) of your prepared slide with the 4x objective below:

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4x, weakest lighting

4x, strongest lighting

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Photo 2 – Activity 1

Insert the image(s) of your prepared slide with the 10x objective below:

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10x, weakest lighting

10x, strongest lighting

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Photo 3 – Activity 1

Insert the image(s) of your prepared slide with the 40x objective below:

40x, weakest light

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40x, strongest light

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10 © 2017 Carolina Biological Supply Company
Post lab Questions – Activity 1

1. Having now completed the lab go back and fill in the “Evidence/Claim statement” and
“Reflection statement” above.

2. Did the image of your slide get darker or lighter as the magnification increased? Why
(answer by defining the field of view and how this will change with
increasing/decreasing magnification)?

As the magnification increased, it became darker. While the photos don't reflect this, it was very
noticeable to me. A field of view is a visible area, in this case the field of view for the
microscope is what we're able to see when looking through the lens. As the magnification
increases, the field of view becomes smaller and smaller as it hones in a more direct area.
Because of this, less light is able to get in through the viewing lens, making the slides appear
darker as we increase the lighting.

Activity 2
1. The 2nd activity for this lab begins on page 12 of your microscope user manual and is
entitled “Brine shrimp” experiment. Please read through pages 12-14 in the microscope
user manual and answer the questions below.

2. IMPORTANT: Do not dispose of your brine shrimp without first doing the following.
Freeze all samples of the brine shrimp overnight and then pour them down the drain.
Brine shrimp are likely not native to your surrounding ecosystem and should therefore be
disposed of properly.

Photo 1 – Activity 2

Insert the image(s) of your drawing(s) of brine shrimp.

I realize this question specifically asked for a drawing, but I assumed we were supposed to insert
a photograph, as that seems to be one of the main points of this lab. Also, I believe you referred
to our “photos” of the brine shrimp in one of the announcements you made, so I hope it's alright
that I've attached a photo rather than a drawing (I'm a horrible artist anyways, so I'd like to think
I'm doing you a favor).

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Unfortunately, my brine shrimp did not hatch. I began to process on the night of 9/29, but have
seen no progress. Instead, this is a picture of some of the eggs I withdrew from the brine solution
they're currently in. Note that the overhead light was used, as I found the bottom illuminator was
not providing very clear photos. This image was taken at 10x magnification.
I'm uncertain why the brine shrimp did not hatch. I made two separate bowls for them,

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one which I began on the night of 9/29, and another I decided to begin midday of 9/30. I was
concerned my first container of brine shrimp eggs had not been done completely correctly, but
didn't want to throw them out or disturb what I had already put together, so I opted to create a
second container and try again. The first container used the exact measurements for water and
salt amounts given in the instructions, and this one saw no results. The second container was
slightly larger, and so I filled it with 2x the water and 2x the salt suggested. I'm not sure if this
played into the eggs inability to hatch. My room has also been fairly cold these last few days, and
I fear the eggs may have not been getting enough light (I work in the evening/night, so they were
left in the dark quite often). I feel the most likely reason they did not hatch is a combination of a
lack of light and heat. I plan to move the containers into my kitchen where they can be in
sunlight more frequently, and hopefully I'll see them hatch in the next couple of days.

Post lab Questions – Activity 2

1. A common microscope lab experiment is to take a printed letter “e” and make a wet
mount slide. Let’s say I take this letter on a slide and place it on the microscope so that it
appears as depicted below (with the bottom of the letter pointing toward the “arm” of the
microscope).

e
When I look at the letter through the eyepiece with the 4x or 10x or 40x objective, would
the imagine of the letter “e” maintain the same orientation? Why or why not? Support
your answer by performing the experiment and place the image that backs up your
answer below.

In this experiment, the e would appear flipped as though it were being displayed in a mirror,
which is to say that it would appear to be flipped horizontally. This occurs because of the
multiple lenses inside of the microscope. The light is refracted through the lenses in a way that
renders the image to the viewer as being flipped horizontally. See the below image for an
example of the letter e observed through the microscope.

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 References

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Carolina Distance Learning. (2017). Fundamentals of Microscopy, Investigation Manual.
Carolina Biological Supply Company.

Clark, M. A., Jung, C., & Douglas, M. (2018). Biology 2e. Houston: OpenStax.

Ward, B. (2021, June 3). Do microscopes invert images? Microscope Clarity. Retrieved from
https://microscopeclarity.com/do-microscopes-invert-images/.

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