LABORATORY NO.

THROWBACK KNOW-HOW ON VIEWING AND

MANIPULATING VARIOUS TYPES OF MICROSCOPE

Introduction  
            A microscope is one of the most important tools utilized by a biologists. It is an
expensive precision instrument that requires special care and handling. As a biology
major students, it is a must that you are equipped with the knowledge and skills on
how to manipulate the microscope. Thus, in this activity, you will recall the major parts
and functions of the microscope, manipulate various types of microscope and learn the
proper care and use of the microscope.

Objectives:
1. Enumerate and identify the parts and functions of the microscope.
2. Compare and contrast images using the different types of microscope.
3. Learn how to properly handle microscope.
4. Calculate magnification of each specimen.

Materials:
Dissecting microscope Petri dish
Compound microscope Cat Flea (whole organism) as provided
Prepared slides Scalpel

 I. Parts, Care and Function of the Microscope

 1. Carrying the microscope

 
a) Always carry the microscope with both hands. Grasp the arm of the microscope
with one hand and place your other hand under the base. (Your instructor will
demonstrate the proper way to transport a microscope.)
b) Always hold the microscope in an upright position (so the eyepiece will not fall
out.)
c) Place the microscope at least 10 cm from the edge, with the arm facing you.

 2.  Microscope Parts – (see separate sheet) Read and make sure you understand the
function and locate each part of the microscope. Be able to classify it into three
different parts.

 Illumination    
a) After you plug the microscope in and turn on the illuminator, rotate the Iris
diaphragm on the underside of the stage in order to adjust the light intensity.
Look through the microscope as you adjust the diaphragm and observe the
changes. Set the diaphragm and move on.
 b) Keeping the light intensity in mind, look through the microscope and move
through the different magnifications slowly. Observe the change in light intensity.
c) Adjust the intensity of light to match the requirements of the objective you are
wishing to view.

Focusing the microscope:

Using a compound microscope:
a) Place a prepared microscope slide, provided to you by your instructor, on the
top of the stage (cover slip up) surface.
b) Position the 4X objective lens into the optical path, making sure that the lens
clicks into position.
c) Look at the microscope with your head to the side. Locate the coarse
adjustment knob which moves the stage up and down. Practice moving the
coarse adjustment knob, seeing how it moves the stage with each turn of the
knob. Using the coarse adjustment knob, move the stage down as far as it
will go. This is the starting position when you first begin to bring an object
into focus. Look into the eyepiece. Slowly, move the stage up by using the
coarse adjustment knob until it comes into focus. Never allow the slide to
come in contact with the objective lens.
d) Turn the fine adjustment knob to bring the specimen into sharp focus.
e) You may need to change the aperture (opening) of the iris diaphragm to
control the brightness. It depends on the specimen and the magnification you
are using.
Moving to a higher magnification:
f) Rotate the revolving nosepiece to position 10X objective into the optical path.
Use ONLY the fine adjustment knob to bring the image into sharp focus.
g) Rotate the revolving nosepiece to position 40X objective into the optical path.
Use ONLY the fine adjustment knob to bring the image into sharp focus.
h) Draw the images seen in each objectives.

 **** NEVER use the coarse adjustment when focusing on the medium and high
power objectives!!!!!

Using a dissecting microscope:

i) Get a sample (whole mount of macro-organism) from your instructor. Place
the sample in the petri dish.
j) Place the specimen on the stage and view it through the eyepiece. Turn
magnification knob until the object is as large as you need it to be.
k) Draw images as seen.

3.  Calculating the Magnification  
 
 1. Observe the numbers etched on the objectives and on the ocular (W10XD). The
TOTAL magnification is found by multiplying the magnification of the objective by
the magnification of the ocular.
 
2. EXAMPLE: If the magnification of the object was DIN 20 and the ocular was
W10XD, the total magnification would be 20 X 10 = 200 X. In other words, the
specimen seen through the microscope would be 200 times larger the viewed
with the naked eye.
 
3. Complete the data table on your answer sheet to find the total magnifications of
each objective on our microscopes.
 
7. Cleaning the lenses  

Lens Paper should be used to clean any of the lenses of the microscope. Lens
paper is provided to you by your instructor. Never use anything other then lens paper
to clean the lens on the microscope, including your fingers.

PARTS AND FUNCTION OF THE MICROSCOPE

Mechanical parts:

a) Body Tube:  hollow cylinder tube that holds the ocular

b) Arm:  Supports the tube and connects it to the base. Serves as the handle of the
microscope.
c) Revolving nosepiece or turret: attached to the base of the body tube and holds
the objectives. Facilitates shifting of the objectives; protected by dust shield that
keeps off the dust from the objective lenses.
d) Stage:  The flat platform where you place your slides.  Stage clips hold the slides
in place.  If your microscope has a mechanical stage, you will be able to move
the slide around by turning two knobs.  One moves it left and right, the other
moves it up and down.
e) Stage aperture: the opening on a microscope slide that allows light to pass
through to the specimen.
f) Base:  The bottom of the microscope, used for support
g) Adjustment knobs: consist of two pairs of knobs which when turned clockwise or
counterclockwise, lower or raise the body tube respectively.
 Coarse adjustment knobs – upper large knob for faster movement of the
body tube when focusing the low power objectives
 Fine adjustment knob – lower smaller knob for slow or little movement of
the body tube when focusing the high power objectives as well as the
 high powered lenses. Also used to get a sharper focus of the object under
the low power objective.

Magnifying parts:
- Eyepiece/Ocular:  detachable short cylinder situated on top of the draw tube
through which one views the specimen; equipped with two set of lenses with
indicated magnifying power; sometimes provided with a pointer used for pointing
specific parts of the specimen.
- Objective Lenses:  Usually you will find 3 or 4 objective lenses on a
microscope.  They almost always consist of:
LPO – low power objectives – 10x
HPO – High power objectives – 40x
Scanner – 4x
OIO- oil immersion objectives – 100x
These objectives when coupled with a 10X (most common) eyepiece lens, we get
total magnifications of 40X (4X times 10X), 100X , 400X and 1000X.  To have
good resolution at 1000X, you will need a relatively sophisticated microscope
with an Abbe condenser.  The shortest lens is the lowest power, the longest one
is the lens with the greatest power.  The high power objective lenses are
retractable (i.e. 40XR).  This means that if they hit a slide, the end of the lens
will push in (spring loaded) thereby protecting the lens and the slide.  All quality
microscopes have achromatic, parcentered, parfocal lenses.
Illuminating parts:
- Illuminator:  A steady light source (110/220 volts) used in place of a mirror. 
If your microscope has a mirror, it is used to reflect light from an external light
source up through the bottom of the stage.
- Condenser Lens:  The purpose of the condenser lens is to focus the light onto
the specimen.  Condenser lenses are most useful at the highest powers (400X
and above).  Microscopes with in stage condenser lenses render a sharper image
than those with no lens (at 400X).  If your microscope has a maximum power of
400X, you will get the maximum benefit by using a condenser lenses rated at
0.65 NA or greater.  0.65 NA condenser lenses may be mounted in the stage and
work quite well.  A big advantage to a stage mounted lens is that there is one
less focusing item to deal with.  If you go to 1000X then you should have a
focusable condenser lens with an N.A. of 1.25 or greater.  Most 1000X
microscopes use 1.25 Abbe condenser lens systems.  The Abbe condenser lens
can be moved up and down.  It is set very close to the slide at 1000X and moved
further away at the lower powers.  
- Diaphragm or Iris:  Many microscopes have a rotating disk under the stage. 
This diaphragm has different sized holes and is used to vary the intensity and
size of the cone of light that is projected upward into the slide.  There is no set
rule regarding which setting to use for a particular power.   Rather, the setting is
a function of the transparency of the specimen, the degree of contrast you desire
and the particular objective lens in use.
How to Focus Your Microscope:  
 The proper way to focus a microscope is to start with the lowest power objective
lens first and while looking from the side, crank the lens down as close to the
specimen as possible without touching it. 
 Now, look through the eyepiece lens and focus upward only until the image is
sharp.  If you can't get it in focus, repeat the process again.  
 Once the image is sharp with the low power lens, you should be able to simply
click in the next power lens and do minor adjustments with the focus knob. 
 If your microscope has a fine focus adjustment, turning it a bit should be all
that's necessary.   Continue with subsequent objective lenses and fine focus each
time.
Name: _______________________ Date Performed: _____________
Teacher Initial: _________________ Date Submitted: _____________

WORKSHEET NO. 1

PARTS OF THE MICROSCOPE

A. Label the parts of a compound microscope below:

Classify the following parts of a compound microscope into mechanical, illuminating and
magnifying parts. Write Mc before the part if it is mechanical, Il if illuminating and Mg if
magnifying.

______________ 1. Mirror ____________6. Draw tube

_______________2. Eyepiece ____________7. Adjustment knobs
_______________3. LPO ____________8.Oil immersion objective
_______________4. Revolving nosepiece ____________ 9. Stage clip
_______________5. Condenser ____________10. Inclination joint
B. Label the parts of a dissecting microscope below:

C. Identify the function of the folowing parts of the microscope:

PARTS: FUNCTIONS:
COMPOUND MICROSCOPE:
1. Objectives
2. Dust Shield
3. Fine adjustment knob
4. Stage clips
5. Coarse adjustment knob
DISSECTING MICROSCOPE:
1. Eyepiece
2. Stage Plate
3. Diopter
4. Eyepiece
5. Stereohead

Answer the following questions:

 1. Compare and contrast compound and dissecting microscope:
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2. What is the proper way to carry a microscope?

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3. What is the purpose of the objectives?
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4. What is the difference between a coarse and fine adjustment knob?

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WORKSHEET NO. 2
 CALCULATING SPECIMENS MAGNIFICATION

INSTRUCTIONS: Drawings should be done in pencil. The circles represent the field of view. Always
remember to record the total magnification each drawings was made in as well as the specimen that was
observed.

COMPOUND MICROSCOPE:

A. B.

SPECIMEN _____________________ SPECIMEN _____________________

MAGNIFICATION: _______________ MAGNIFICATION __________________

C.

SPECIMEN _____________________
MAGNIFICATION: _______________

DISSECTING MICROSCOPE:
E. F.

SPECIMEN _____________________ SPECIMEN _____________________

MAGNIFICATION: _______________ MAGNIFICATION __________________
Answer the following questions:

1. What happens to the specimen when viewed from 10x to 40x?

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2. What is the total magnification power of your microscope under the following
operating conditions?

OBJECTIVE OCULAR TOTAL MAGNIFICATION (X)

4x 10 x _______________
10x 10 x _______________
40x 10 x _______________

3. Differentiate the images seen in compound and dissecting microscopes.

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Conclusion