FACULTY OF SCIENCE

BSc (Hons) Biotechnology

May 2014 Trimester
LABORATORY REPORT
Unit Code & Title: UDBB1104 Cell Biology

Practical Group: P1

Experiment Title: The Compound Microscope

Date of Submission: 24/06/14
Instructor: Dr Kunasundari a/p Balakrishnan
Declaration
I certify that this report is of my own work and does not contain any
unacknowledged work from any other sources.
No. Name
1 Eswaran

Student ID
1401660

Signature

Objectives:
1.
2.
3.
4.
5.

To learn about the correct method of using a compound light microscope.

To be able to recognize the function of the parts of a compound microscope.
Able to make a wet mount
To utilize the laboratory skills learned to explore the microscopic world
Learn how to care for a light microscope

Introduction:

A compound microscope is an instrument used to see objects that are far too small for the
human eye. The compound microscope is a light microscope since it uses visible light as the source
of illumination. A compound microscope operates with a series of lenses which forms a clearly
focused that is many times larger than the actual specimen itself (magnification).
This magnification is achieved when light rays from an illuminator passes through a
condenser, which has lenses that direct the light rays through the specimen. This point onwards,
the light rays pass into the objective lenses. The image of the specimen is magnified again by the
ocular lens, or eyepiece. Most compound microscopes used these days have several objective
lenses, including 10x (low power), 40x (high power), and 100x (oil immersion). Most ocular lenses
magnify specimens by a factor of 10. By multiplying the magnification of a specific objective lens
with that of the ocular lens, we may conclude that the total magnifications would be 100x for low
power, 400x for high power, and 1000x for oil immersion. However, some compound light
microscopes can achieve a total magnification of 2000x with the oil immersion lens.
Resolution (also known as the resolving power) is the ability of the lenses to distinguish
fine detail and structure. This specifically refers to the ability of the lenses to distinguish two points
a specified distance apart. In general the principle of microscopy is that the shorter the wavelength
of light used in the instrument, the greater the resolution of the image formed would be. The white
light used in a compound light microscope has a relatively long wavelength and cannot resolve
structures smaller than about 0.2 m. This factor limits the magnification achieved by a compound
light microscopes to about 2000x.
In order to obtain a sharp and finely detailed image under a compound light microscope,
specimens must be made to contrast sharply with their medium. To attain such contrast, we must
change the refractive index of specimens from that of their medium (refractive index is a measure
of the light-bending ability of a medium). We may change the refractive index of specimens by
staining them. (Gerald J. Tortora, 2012)

Materials:
1.
2.
3.
4.
5.
6.
7.
8.

Compound Light Microscope

Prepared slide of letter D
Prepared slide of colour fiber
Prepared slide of Spirogyra
Colour fibers
Glass slides
Cover slips
Water

Method:

1. Before using the compound microscope, the objective lens of the microscope is set to the
lowest resolution (10x). The compound microscope is plugged into a power supply and
switched on.
2. A prepared slide of a colour fiber is observed under the microscope using the lowest
magnification and the slowly progress to the higher magnification. The details of the fiber
is observed.
3. Next, a prepared slide of the letter D is observed under the microscope. The size and
orientation of the letter is observed and recorded.
4. The slide is then moved in multiple direction and direction the microscopic image moves
in response is recorded.
5. A fiber which has been dyed with two colours is placed on a glass slide, a drop of water
is then added to the fiber and a plastic cover slip is placed on top of the fiber.
6. This slide is the observed under the microscope and the different coloured fibers are
identified.
7. The diaphragm is then closed until the light is almost cut off and the fiber is observed
again.
8. A prepared slide of spirogyra is then observed microscopically.
9. Once the image is in focus the fine adjustment knob is raised until the surface of the cell
is visible, the position of the fine adjustment knob is recorded.
10. Later, the fine adjustment knob is lowered until the lower limit of the cell is visible, the
position of the fine adjustment knob is again recorded.
11. The spaces moved by the fine adjustment knob is identified and the thickness of the cell
is recorded.

Results:

(Figure 1.1: A single fiber observed under the microscope)

Observation:
A dark toned fiber is seen against the bright field background. Tiny strands branching out from the
fiber are also visible

(Figure 1.2: A printed material, the letter D is observed under the microscope)
Observation:
The letter D seems to be inverted upside down
4

Yellow strand of
fiber

Orange strand of
fiber

(Figure 1.3: A fiber which has been dyed with different colours observed under the microscope)
Observation:
A single fiber which has been dyed with different colours are observed under the microscope and
the strands with different colours are visible through the microscope. This particular fiber has
strands dyed orange and yellow.

(Figure 1.4: Spirogyra observed under the microscope)

Observation:
The spirogyra appears to be layered one above another. This could be proven when the fine
adjustment knob is rotated and the spirogyra on top loses focus and the spirogyra below gains
focus.
5

Discussion:

1.
Knowing that the low-power field has a diameter of 1.6 mm, calculate the area of the field
by using the formula Area = r2. The area of the low-power field of my microscope is____mm2.
The area of the high-power field of my microscope is ____mm2.

2.
The size of the letter D is magnified 400x (including the magnification of the ocular lens)
of its original size. The image of the letter D which is seen through the microscope seem to be
inverted.
The image observed through the compound microscope is inverted due to the effect of the
lenses it is made of. The objective lens in a compound microscope has a relatively short focal
length. When the light passes through a specimen, passing through the objective lens, and the focal
point of the objective lens, the image formed would be inverted. This inverted image is the object
which is seen by the ocular lens and the eyepiece lens role is to magnify and enlarge the image
created by the objective lens. Due to this, the image that is seen when looking through a compound
microscope is inverted when compared to the real specimen examined. (McDoogleburger, 2013)
When the slide is moved in one direction, the image that is seen when looking through a
compound microscope seem to move in the opposite direction. This happens because the image
seen through the microscope is inverted therefore so does the direction the image observed move.
For instance if the slide is moved to right, the image seen through the microscope would move left.
Same goes to all the other directions.

3.
A fiber dyed in two colours are observed under a compound microscope, when the
diaphragm is opened wide, we are able to see both the orange strand and the yellow strand clearly.
It is also easy to differentiate these two colours. Both the orange and yellow dye on the fabric is
observable because the well illuminated background creates a good contrast against the dye colour
of the fabric.
However when the diaphragm is closed until the light is almost cut off, it is difficult to
differentiate the orange and yellow strands in the fiber. This is because the closed diaphragm
blocks the light from the condenser lens to pass through the specimen. This causes a dark
background to be formed which is not in contrast with the fiber to be observed. Without a good
illumination, the details in the fiber are not visible to the observers eyes. From this observation
we may conclude that the compound microscope is a bright-field microscopy and light plays a
vital role in creating contrast in the images viewed through the compound microscope.

4.
In order to estimate the thickness of a spirogyra the reading of the fine adjustment knob
which makes out the upper surface of the cell and the reading of the fine adjustment knob which
makes out the lower limit of the cell must be identified
Based on the results:
Reading of the fine adjustment knob which makes out the upper surface of the cell = 53
Reading of the fine adjustment knob which makes out the lower limit of the cell = 71
Change in spaces on the fine adjustment knob = Reading of the fine adjustment knob which makes
out the lower limit of the cell - Reading of the fine
adjustment knob which makes out the upper
surface of the cell
= 71 53
= 18 spaces
From the lab manual it is known that, 1 space movement on the fine adjustment knob is equal to
0.002mm or 2m
Therefore we may conclude that,
Thickness of a spirogyra cell = Change in spaces on the fine adjustment knob x 2m
= 18 x 2m
= 36m
The estimated thickness of a spirogyra cell is 36m.

Conclusion:
The compound microscope is an important and efficient instrument in viewing materials
which are impossible to be seen by the naked eyes. Whereby, factors such as magnification,
resolution and contrast determines the quality of the image produced by the compound microscope.
By calculating the thickness of a spirogyra cell the compound microscope is also proven to be an
instrument of measurement. From this experiment may conclude that the compound microscope
is an essential instrument to explore the microbial world.

References
Gerald J. Tortora, B. R. F. C. L. C., 2012. Microbiology An Introduction 11th edition pg 55-57. Paris:
Pearson.

McDoogleburger, A., 2013. Why Do Compound Microscopes Invert the Images?. [Online]
Available at: http://www.ehow.com/how-does_5208588_do-compound-microscopes-invertimages_.html?ref=Track2&utm_source=ask
[Accessed 22 June 2014].