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Max, Tom, John, Creasic, Leo,

Using Compound Microscopes


15/10/2016
On my honor, I have neither received nor
given any unauthorized aid on this
assignment.

INTRODUCING
What is a compound microscope? Quoting wisegeek.org, A compound microscope is
a microscope fitted with two or more convex lenses. The high magnification produced by
these lenses together enables a detailed study of micro-organisms, cells and tissues. These
types of microscopes are therefore widely used in scientific and medical research.
According to history-of-the-microscope.org, The first compound microscope was invented
by Zacharias Jansen and his father Hans in 1590s. They put several lenses together in a tube
and discover the compound microscope. Their first microscope was not that good because the
maximum magnification was only about 9x and it was still not that clear.
When should we use the compound microscope? Quoting kids.britannica.com, The
compound microscope uses a lens called the objective to produce a primary magnified image
and another called the eyepiece or ocular to magnify this image. In practice, both objective
and eyepiece are actually composed of several lenses to neutralize the optical defects inherent
in lenses made of a single kind of glass. According to microscope-detective.com, There are
2 main microscopes, Optical Microscopes and Electron Microscopes. Optical Microscopes
also known as light microscopes, it means that it use light. There are three types of Optical
Microscopes and they are Compound Microscopes, Stereo Microscopes, and Confocal
Microscopes. I already explain how Compound Microscope works already so I will not
explain it again. A Stereo Microscope is different from a Compound Microscope in a few
ways. A Stereo Microscope has 2 eyepieces and Compound Microscope only have one. It has
2 eyepieces so it can produce a three-dimensional images and that is the main thing about
Stereo Microscope. Now comes Confocal Microscopes, the light of the Confocal
Microscopes comes from a laser. It scans the sample and sends it to the computer and there
are no eyepieces. Electron Microscopes are another type of microscope. This kind of
microscope scans with electrons and it will give you a clearer image. There are two main type
of Electron Microscopes and they are Scanning Electron Microscope (SEM) and
Transmission Electron Microscope (TEM). A SEM sends a beam of focus electrons to the
sample and then it will bounce off and creates a three-dimensional surface image. A TEM
also send a beam of electron to a thin sample and then create a two-dimensional image. There
are also others microscope to like Scanning Probe Microscope, Scanning Acoustic
Microscope, X-ray Microscope. A Scanning Probe Microscope scans a sample with a
physical probe. Scanning Acoustic Microscope use sonar or sound waves to identify the
sample. X-ray Microscope takes an x-rays to produce the image of the sample.

MATERIAL
Distilled water
Methylene blue
Onion
Onion root
Yogurt (undiluted)
Yogurt (diluted)
Compound Microscope
Toothpick
Gloves
Glass Slides
Coverslips
Pipette
Fire
Check
Oil Filter

PROCEDURE
Onion Cells
1) Use a knife to cut the onion slightly.
2) Put it in a rectangle glass and covers it with a square plastic and cover it up.
3) Then we put it in the microscope to see it.
4) First we look at 4x.
5) Move the slide to make the sample right in the middle of the light to let us see it
better.
6) Move the slide slowly up or down until you see it.
7) Now we start again with step four then go down but now with 10x.
8) Observe and take pictures of 10x and 4x.
Onion Root Cells
1) Cut the root out of the onion and then cut it in smaller size.
2) We put it in a rectangle glass and cover it with a square plastic and cover it up.

3) We put it in the microscope to see it.


4) First we look at 4x.
5) Move the slide to make the sample right in the middle of the light to let us see it
better.
6) Move the slide slowly up or down until you see it.
7) Now we start again with step four then go down but now with 10x.
8) Observe and take pictures of 10x and 4x.
Check Cell
1) Wear Gloves
2) Take a toothpick and then scrape slightly to get the check cell.
3) After that we put it in a rectangle glass.
4) Give it to Mr Bill.
5) Mr Bill put methylene blue to it to let us see the check cell easier.
6) We then cover it with a square plastic.
7) We put it in the microscope to see it.
8) We start at 4x.
9) Move the slide to make the sample right in the middle of the light to let us see it
better.
10) Move the slide slowly up or down until you see it.
11) Now we start again with step eight then go down but now with 10x.
12) Observe and take pictures of 10x and 4x.
Yogurt (undiluted)
1) Wear gloves
2) Take the toothpick and then put it in the yogurt and take it out
3) Put it in the glass
4) Let it dry for 3 minutes
5) Held over fire a few time.
6) Give Mr Bill to let him put the methylene blue
7) We then put the plastic to cover it
8) We put it in the microscope to see it.
9) We start at 4x.

10) Move the slide to make the sample right in the middle of the light to let us see it
better.
11) Move the slide slowly up or down until you see it.
12) Now we start again with step nine then go down but now with 10x.
13) Observe and take pictures of 10x and 4x.
Yogurt (diluted)
1) Wear gloves
2) Take the toothpick and then put it in the yogurt and take it out
3) Put it in the glass
4) Let it dry for 3 minutes
5) Held over fire a few time.
6) Give Mr Bill to let him put the methylene blue
7) We then put the plastic to cover it
8) We put it in the microscope to see it.
9) We start at 4x.
10) Move the slide to make the sample right in the middle of the light to let us see it
better.
11) Move the slide slowly up or down until you see it.
12) Now we start again with step nine then go down but now with 10x.
13) Observe and take pictures of 10x and 4x.
DATE & RESULTS

(Onion Cells in 4X)


In this picture I see a pink, purple onion cells with dots in it.

(Onion Cells in 10X)

In this picture you can see the colour is fading and the dots is getting bigger because we are
getting closer.

(Onion Root Cells in 4X)


You can see the green colour of the root and string in the middle. You can also see little dot
out the corner.

(Onion Root Cells in 10X)


You can see the colour change from green to brown and there are also dots in here but in 4X
you cannot see it.

[Yogurt (diluted) in 4X]


The yogurt is yellow with different size. There is also bacteria too.

[Yogurt (undiluted) in 10X]


Now we are looking at the yogurt and it does not have anything except a little methylene
blue.

[Yogurt (undiluted) in 4X]


In this diluted yogurt it contains bacteria and a lot of yogurt.

[Yogurt (undiluted) in 10X]


In this picture you can see the bacteria is getting many and the yogurt is lesser.
DISCUSSION

(Onion Cells)
Compering this picture to our picture you can see the colour is the same but the structure is
different. In this image the structure is like a brick wall that did not get paint and my picture
is like a wall that has been painted but it gat holes in it.

(Onion Root Cells)


Compering this one and ours you will notice that this one got a different colour and it is
maybe 40x or 100x (I am just guessing) and ours is only 4x and 10x

(Yogurt)
Compering this and our picture you can see it is the same but different colour they have a lot
of bacteria and some yogurt like us. So what bacteria are in yogurt? According to Elizabeth
Brown, in yogurt there are three kinds of bacteria and they are Lactobacillus,
Bifidobacterium, and Streptococcus Thermophilus. Lactobacillus is a name for one type, or
genus, of bacteria. It is naturally found in the digestive system and urinary tracts, and also in
foods. Most of yogurt is made from Lactobacillus. Bifidobacterium is a good bacteria found
in a healthy gastrointestinal, it is also found in fermented foods too. Streptococcus
Thermophilus have 81 type of it are found in yogurt or cheese. Quoting microbewiki, The
circular chromosome of Bifidobacterium longum has a genome approximately 2,260,000 bp
in length with a 60% G-C content. More research is being done on sequencing other species
of Bifidobacterium, especially to understand its probiotic qualities. Quoting
sites.google.com, All Lactobacilli are physically rod-shaped, and often form in chains or
pairs. They form straight chains, varying in length, which are called Strepto chains. They can
also form into round and spiral shaped bacteria under certain circumstances. Quoting
microbewiki, Streptococcus thermophilus is a gram-positive bacterium; the cell wall is
composed of N-acetylglucosamine (NAG) and N-acetylmuranic acid (NAM), which is bond
by ether bonds. This unique structure allows S. thermophilus to endure elevated temperatures,
which is useful for much industrial dairy fermentation requiring the process of milk at higher
temperatures. It is non-motile, consists of coccus (circular) cell structure, produces no spores,
and lives in pairs or chains (size of chain varies). S. thermophilus is a facultative anaerobe,
which is an organism that is capable of generating ATP through both aerobic respiration and
fermentation (depending on the presence or absence of oxygen). Furthermore, S.
thermophilus lacks genes or contain pseudogenes expressing surface protein (excluding
lipoproteins); pathogenic streptococci use these surface proteins to adhere to mucosal
surfaces and evade host defense mechanisms. [Surface protein such as sortase-anchored
surface proteins, an important virulence factor of pathogenic streptococci, is not present on
the surface of S. thermophilus.
By doing this I learned how to do a lap report because I have never done one before
and this is the first time. I also learned that lab is fun but you need to be really careful and
wear safety equipment in the lab. You also need to hear what the teacher says too. I could
improve my experiment by taking more pictures.
RESEARCH QUESTIONS

The purpose of methylene blue is to make the sample easier to see. When you use methylene
blue you need to wear gloves because it is very poisonous.
Quoting flinnsci, The upper limit of the resolving power of light microscopes is slightly
above 1000X. Objectives of 90 to 100X, when coupled with a 10X eyepiece, approach that
upper limit. Even in the range of 900 to 1000X, a clear image is only possible if every bit of
available light is directed through the microscope optics to the viewer's eye. Immersion oils
play an essential role in maximizing the amount of light producing the image the viewer sees.
In the airspace between the slide and the objective lens, light is refracted, scattered, and
effectively lost. This happens because the refractive index of air (approximately 1.0) is very
different from that of glass (approximately 1.5), and light passing through a glass/air interface
is refracted (bent) to a large degree. By reducing the amount of refraction at this point, more
of the light passing through the slide will be directed to the very narrow diameter lens of the
high-power objective. The more light, the clearer the image. Placing a material with a
refractive index equal to that of glass in the airspace between slide and objective directs more
light through the objective and produces a clearer image. Immersion oils are formulated for
just this purpose. High-power objectives of 90X or higher are almost invariably intended for
use with oil and will be engraved with the words, "oil", or "immersion", or "HI"
(homogeneous immersion). These objectives are assembled with special sealants that prevent
penetration of oil into the lens system. Applying oil to an objective not designed for
immersion will ruin the objective. Immersion oils are commonly available in two viscositieslow viscosity (Type A), and high viscosity (Type B)-and should be labeled with a refractive
index of 1.515. The low viscosity oil is applied to the airspace between slide and objective,
the high viscosity oil is (less commonly) applied between the condenser and the slide. Low
viscosity oil between slide and objective:

1. With low- or medium-power objective, locate a point or area of interest on the slide and
center it in the image field.

2. Rotate the objective turret so that the high power objective is just to one side of the slide.
Place a single drop of immersion oil (low viscosity, Type A) on the slide (using the circle of
light from below as a guide) and place a drop directly on the objective lens. Failure to apply
oil to the objective will likely result in trapped air and reduced image quality.

3. Slowly rotate the high power objective into place and adjust the fine focus to fully resolve
the image.

High viscosity oil between condenser and slide (optional). Condensers with a numerical
aperture (N.A.) of 1.0 and greater (usually engraved directly on the condenser) are also sealed
to prevent oil penetration. Do not immerse condensers with an N.A. less than 1.0.
1. Before placing the slide on the microscope stage, rack the condenser down (using the
condenser focusing mechanism) and apply a drop of oil (high viscosity, Type B) to the
condenser lens.
2. Apply a drop of oil to the bottom of the slide directly below the specimen, and place the
slide on the stage so that the drops will meet when the condenser is raised.
3. Raise the condenser until the drops converge. Follow the steps detailed above to oil the
slide to the objective.

Work Cited
"microscope." Compton's by Britannica. Britannica Online for Kids.
Encyclopdia Britannica, Inc., 2016. Web. 15 Oct. 2016.
<http://kids.britannica.com/comptons/article-204628/microscope>.
"Who Invented the Microscope? A Complete Microscope History." A Complete Microscope
History. N.p., n.d. Web. 15 Oct. 2016. <http://www.history-of-the-microscope.org/history-ofthe-microscope-who-invented-the-microscope.php>.
"What Is a Compound Microscope?" WiseGEEK. N.p., n.d. Web. 15 Oct. 2016.
<http://www.wisegeek.org/what-is-a-compound-microscope.htm>.
"Types of Microscopes and Their Uses." Types of Microscopes and Their Uses. N.p., n.d.
Web. 15 Oct. 2016. < http://www.microscope-detective.com/types-of-microscopes.html>.
"File:Red Onion Cells.JPG." Wikipidia. N.p., n.d. Web.
<https://en.wikipedia.org/wiki/File:Red_Onion_Cells.JPG>.
Hale, Kristi. "Onion Root Tip 2.jpg." Santiago Canyon Collage. N.p., 3 Sept. 2011. Web.
<http://www.sccollege.edu/departments/biology/biology%20109%20lab%20review%20for%
20practicum%201/forms/thumbnails.aspx>.
BROWN, ELIZABETH. "List of Good Bacteria in Yogurt." LIVESTRONG. N.p., 9 June
2015. Web. <http://www.livestrong.com/article/349067-list-of-good-bacteria-in-yogurt/>.
"Bifidobacterium." MicrobeWiki. N.p., 23 July 2010. Web.
<https://microbewiki.kenyon.edu/index.php/Bifidobacterium>.
"Structure - Lactobacillus Bulgaricus Bacteria Site." Structure - Lactobacillus Bulgaricus
Bacteria Site. N.p., n.d. Web. 16 Oct. 2016.
<https://sites.google.com/site/lactobacillusbulgaricussite/structure>.
"Streptococcus Thermophilus." Microbewiki. N.p., n.d. Web.
<https://microbewiki.kenyon.edu/index.php/Streptococcus_thermophilus>.
"Teacher Resources." What Is Immersion Oil Used for and What Is the Proper Technique for
Using It? N.p., n.d. Web. 16 Oct. 2016. <https://www.flinnsci.com/teacherresources/biology/frequently-asked-biology-questions/general-questions/what-is-immersionoil-used-for-and-what-is-the-proper-technique-for-using-it/>.