Using Compound Microscopes

Group:
Alice, Charice, Holly, Lala, Janet
October 14, 2016
INTRODUCTION:
According to HISTORY of the Microscope, Zacharias Jansen and his father
Hans invented the first compound microscope in 1595. They started with experiment
these early magnifier lenses, called flea glasses. They put many of these glasses into a
tube. Jansen and his father found out that the object on the end of the tube appeared
larger than normal. But the maximum magnification of their first compound
microscope was only around 9x and the images were blurry (Who invented the
microscope? A complete Microscope History).

The first microscope found by Zacharias Jansen and his father Hans. (Source:
history-of-the-microscope.org)
Compound microscopes are usually found in science lab. Compound
microscopes are electronic devices that use to focus on a species to the cells
inside it. There are many different types of microscopes. Quoting
MicrobeHunter Microscopy Magazine, Optical Microscopes: These
microscopes use visible light (or UV light in the case of fluorescence
microscopy) to make an image. Optical microscopes can be further subdivided
into several categories:
Compound Microscope: These microscopes are composed of two
lens systems, an objective and an ocular (eye piece).
Stereo Microscope: They are useful for observing opaque objects.
Confocal Laser scanning microscope: These devices are reserved for
research organizations. They are able to scan a sample also in depth. A
computer is then able to assemble the data to make a 3D image.
X-ray Microscope: As the name suggests, these microscopes use a beam of xrays to create an image.
Scanning acoustic microscope: These devices use focused sound waves to
generate an image. They are used in materials science to detect small cracks or
tensions in materials.
Scanning Helium Ion Microscope: these devices use a beam of Helium ions
to generate an image.
Neutron Microscope: These microscopes are still in an experimental stage.
Electron Microscopes:
Transmission electron microscopy: In this case, the electron beam is
passed through the sample. The result is a two dimensional image.
Scanning electron microscopy: The electrons do not go through the
sample but bounce off. It is possible to visualize the surface structure
of the specimen. The image appears 3 dimensional.
Scanning Probe Microscopes: It is possible to visualize individual atoms
with these microscopes (Different Types of Microscopes).

MATERIALS:
Compound microscope
Glass slides
Coverslips
Onion
Onion root
Yogurt (diluted)

Yogurt (undiluted)
Methylene blue
Cheek
Toothpicks
Gloves
Pipette
Distilled water
Fire
Oil (to build fire)

PROCEDURE:
- For all samples, prepare glass slides and cover slips.
Onion Cells:
1. Take a small knife and cut a very slight piece of onion to fit between the glass
slide and the cover slip.
2. Put the piece onto the glass slide and add the coverslip.
3. Place the onion slide on the compound microscopes stage and use the stage
clip to hold the slide.
4. First, use the magnification of 4x, the low-power objective, to focus on the
onion.
5. Move the slide slowly until you see the slide of onion.
6. Use the fine adjustment knob to focus the area.
7. Repeat from step 4 with the magnification of 10x, medium-power objective.
8. Finally, take pictures and observe.
Onion Root:
1. Just like onion cells, take a small knife and cut a slight piece of onion root.
2. Put it on the glass slide and cover with the slip.
3. Place the slide on the stage of compound microscope and hold the slide in
place by the stage clip.
4. Use the 4x objective to focus on the onion root slide.
5. Move the stage in different directions to see the onion root.
6. Use the fine adjustment knob to focus on the onion root.
7. Repeat from step 4 with 10x objective.
8. Finally, take pictures and observe.
Cheek Cells:
1. Put on the gloves.
2. Take a toothpick and scrape the cheek inside your mouth.
3. Put the cheek cell on the glass slide.
4. Wait for 3 minutes for the cheek to dry.
5. Hold on fire for 3 to 4 times.
6. Then, use the pipette to drop methylene blue on the cheek cell slide and put on
the coverslip. (Use distilled water to clean up if the methylene blue mess
outside)
7. Place the slide on the compound microscopes stage and use the stage clip to
hold it in place.
8. Use the 4x objective first and move the stage in different directions to find the
cells.
9. Use fine adjustment knob to see the cells clearly.

10. Repeat from step 7 with the magnification of 10x.

11. Take pictures and observe.
Yogurt (diluted):
1. Put on the gloves.
2. Dip a toothpick into the yogurt and take a small amount.
3. Spread it on the glass slide and wait for 3 minutes for the yogurt to dry.
4. Hold the slide over the fire for 3 to 4 times.
5. Use the pipette to put the methylene blue on the sample.
6. Take the coverslip and put on the slide. (Use distilled water to clean up if the
methylene blue mess outside)
7. Put the slide on the stage and use the stage clip to hold.
8. Start with 4x objective, use different knobs to move the stage until you see the
cells and use the fine adjustment knob to see it clearer.
9. Repeat step 8 with 10x objective.
10. Take pictures and observe.
Yogurt (undiluted):
1. First, we put on the gloves.
2. Take a toothpick and dip it in the yogurt.
3. Take out a small amount and spread it on the glass slide.
4. Wait for 3 minutes to dry and put it over the fire 3 to 4 times.
5. Drop the methylene blue on the sample with a pipette and place the coverslip
on top of the sample. (Use distilled water to clean up if the methylene blue
mess outside)
6. Put the slide on the compound microscopes stage and hold it in place by the
stage clip.
7. Use 4x objective and slowly move the stage until you see the cells.
8. Use fine adjustment knob to see the cells clearly.
9. Repeat from step 8 with 10x objective.
10. Take pictures and observe.
DATA & RESULTS:

- Onion Cells (4x objective)

- Onion Cells (10x objective)

Onion Cells:
In 4x objective, I see purple color and it looks like a net. It has many square
bricks inside.
In 10x objective, it also looks like a net but I see a darker color of purple and
brown in it. And I see that the square bricks are stacking on each other.

- Onion Root (4x objective)

- Onion Root (10x objective)

Onion Root:
In 4x objective, I see green and clear color. It has some cross lines and little
dots in it.
In 10x objective, the onion root is also green and clear. It also has cross lines.
It looks like the enlarge version of the onion root in 4x objective.

- Cheek Cells (4x objective)

- Cheek Cells (10x objective)

Cheek Cells:
In 4x objective, I see many blue dots, which is a nucleus. The image looks like
a planet.

In 10x objective, I see the image with dark blue nucleus.

Our team cannot see anything in 40x and 100x objectives.

- Diluted Yogurt (4x objective)

- Diluted Yogurt (10x objective)

Yogurt (Diluted):
In 4x objective, it has an almost-triangle shape with blue border.
In 10x objective, it looks like blue crystal.
Our team cannot see anything in 40x and 100x objectives.

- Undiluted Yogurt (4x objective)

Yogurt (Undiluted):
In 4x objective, it looks like it is being scratch. I can see blue and white
scratch line.
Our team cannot see anything in 10x, 40x, and 100x objectives.
DISCUSSION:
Online Microscopic Pictures:

- Onion Cells
(Source: amrita.olabs.edu.in)

- Onion Root Cells

(Source: Google Image)

According to the picture of amrita.olabs.edu.in, onion includes onion cells,

cell wall, nucleus, cytoplasm, and vacuole. The nucleus appears at the side of
cytoplasm. The vacuole is in the center of the cell, surround by cytoplasm (Onion and
Cheek Cells).
Quoting vlab.amrita.edu, The most commonly used root tips in labs to study
mitosis are onion, wheat, lentil, barley and alfalfa. An onion root tip is a rapidly
growing part of the onion and thus many cells will be in different stages of mitosis.
The onion root tips can be prepared and squashed in a way that allows them to be
flattened on a microscopic slide, so that the chromosomes of individual cells can be
observed easily. The super coiled chromosomes during different stages of mitosis
present in the onion root tip cells can be visualized by treating with DNA specific
stains, like Feulgen stain and Acetocarmine stain (Mitosis In Onion Root Tips).
According to LIVESTRONG.COM, bacteria that are commonly found in
yogurt are lactic acid bacteria. These bacteria are good bacteria, also known as
probiotics. Lactic acid bacteria are including lactobacillus, bifidobacterium, and
streptococcus thermophilus (Elizabeth Brown).

- Lactobacillus (Source: Google Image) - Bifidobacterium (Source: Google Image)

- Streptococcus Thermophilus (Source: Google Image)

- Cheek Cells (Source: Google Image)

Quoting Reference, Cheek cells are generally irregularly shaped and are
always flat cells. The cells are made up of many parts including a very thin membrane
on the outer part of the cell (What is the general shape of a cheek cell?).

Compare:
My microscopic picture of onion cells has the same bricks, which is the cell
wall, as the online microscopic picture. My picture cannot see the nucleus and
cytoplasm.
My picture of onion root is completely different from the online picture. The
online picture has many dots. It has rectangle bricks and some of the bricks do
not have any dot.
My picture of cheek cells has the nucleus just like the online picture. But my
microscopic picture does not have the smaller dots around the nucleus and the
bigger dot that contain the nucleus inside.
My picture of yogurt (diluted and undiluted) is totally different from the
online picture. My picture does not have any of the bacteria that should have
in yogurt (lactobacillus, bifidobacterium, or streptococcus thermophilus).

Through this experiment, I have learn how to use compound microscope, how to
control it. I learned that we should always start at 4x objective when using the
compound microscope. When we use 100x objective, we must put oil on the slide in
order to see. I learned that we should lab safety equipment when we entered the lab.
And we should use gloves when we do something that relate to the methylene blue.

RESEARCH QUESTIONS:
According to IHCWORLD, we must wear gloves while working with
methylene blue. Avoid eye contact and skin contact. Dont inhale or ingest. Keep
away from heat, such as fire, alkalis, dichromates, iodides, and reducing agents.
Remember to wash hands after working with methylene blue (Methylene Blue).
Quoting IHCWORLD, Methylene blue is a known animal mutagen and should not be
handled by staff who are pregnant. It is an eye irritant and may irritate skin. It is
moderately to highly toxic by oral and intravenous routes. Eye contact can cause
staining of the eye, stinging and lacrimation. Inhalation may cause Heinz bodies
haemolytic anaemia, methaemoglobinaemia and cyanosis. Ingestion may cause
dryness of mouth, flushed skin, rapid pulse, blurred vision, dizziness, haemolytic
anaemia, cyanosis and mehaemaglobinaemia. The powder is combustible and should
never be handled close to heat or a naked flame (Methylene Blue).
According to enotes, the purpose of methylene blue in preparation of human
cheek cells and yogurt slides is that it helps the DNA in the nucleus to be seen
through the microscope light. It causes the nucleus to stand out. If there is no
methylene blue, the cells will be hard to see (What is the Purpose of Adding
Methylene Blue to a Specimen?)
Quoting MICROSCOPEGENIUS.COM, By using a relatively thick oil,
usually cedar wood oil, light is kept from scattering as much, allowing more of it to
maintain the correct angle to enter the lens, and thus your sample becomes viewable
and easier to focus on (MICROSCOPE 101: OIL IMMERSION LENS
TECHNIQUE).
To use 100x oil immersion, we must use 4x objective and 10x objective first
to know the exact area. Then, drop a little amount of oil on the coverslip, not on the
sample, and turn to 100x objective. While using 100x objective, we only use the fine
adjustment knob and turn it very slow and look at it carefully until you see the image.

Works Cited
Who Invented the Microscope? A Complete Microscope History. A Complete
Microscope History,
http://www.history-of-the-microscope.org/history-of-the-microscope-whoinvented-the-microscope.php
Different Types of Microscopes. MicrobeHunter Microscopy Magazine, 31 Dec.
2015,
http://www.microbehunter.com/different-types-of-microscopes/
Our Objective. Onion and Cheek Cells (Theory) : Class 9 : Biology : Amrita Online
Lab,
http://amrita.olabs.edu.in/?sub=79&brch=15&sim=125&cnt=1
Brown, Elizabeth. List of Good Bacteria in Yogurt. LIVESTRONG.COM,
LIVESTRONG.COM, 9 June 2015,
http://www.livestrong.com/article/349067-list-of-good-bacteria-in-yogurt/

What Is the General Shape of a Cheek Cell? Reference,

https://www.reference.com/science/general-shape-cheek-cell48e19b794260ed17#
Objectives: Mitosis in Onion Root Tips (Theory) : Cell Biology Virtual Lab II :
Biotechnology and Biomedical Engineering : Amrita Vishwa Vidyapeetham
Virtual Lab,
http://vlab.amrita.edu/?sub=3&brch=188&sim=1102&cnt=1
ABC of Safety in the Biological Sciences - Methylene Blue. ABC of Safety in the
Biological Sciences - Methylene Blue,
http://www.ihcworld.com/royellis/abcsafe/chemicals/methylene-blue.htm
What Is The Purpose Of Adding Methylene Blue To The Specimen. Enotes.com,
Enotes.com,
http://www.enotes.com/homework-help/want-ask-about-methylene-blue-whatpurpose-using-347231
Microscope 101: Oil Immersion Technique Explained (How To Guide).
MicroscopeGeniuscom, 3 Sept. 2014,
http://microscopegenius.com/microscope-101-oil-immersion-lens-technique/
Honor Code: On my honor, I have neither received nor given any authorized
aid on this assignment.