THE MICROSCOPE

OBJECTIVES
After completing this exercise, you should be able to:
1. Identify the parts of a compound microscope and described
the function of each.
2. Describe and demonstrate the correct way to (a) carry a
microscope, (b) clean the lenses, (c) focus with each
objective, and (d) calculate the total magnification.
3. Define all terms in bold print.
The effective use of a compound
microscope is essential for the study of
cells and tissues. This exercise will introduce
you to the parts, care, and use of a
compound microscope.
PARTS AND MECHANISMS- The parts of a
compound microscope can be conveniently divided
under three headings. The mechanical parts hold all
structures together or control their use while the
optical and illuminating parts regulate the light rays
including the formation of images.
I. MECHANICAL PARTS

1. Base – It supports the whole microscope.

The base is the bottom portion of the microscope.
It contains the lamp and the on/off switch.
Ideally, the lamp should have a voltage control to
vary the intensity of light. The lowest possible voltage
that provides a good image should be used to
extend the life of the lamp.
2. Pillar – This portion extends upwards from the
base supporting all other parts.

3. Inclination Hinge or Joint – The inclination

joint allows tilting of the microscope at any
desired angle, often as a matter of comfort.

4. Stage –The stage is the platform on which a

microscope slide is placed for viewing. It
contains the specimens to be examined.
5. Stage Aperture – A central opening of the stage, it allows
the transmission of light rays from below.

6. Spring Clips – Usually two, they hold the slide securely in

position.

7. Arm –The arm rises from the base and supports the rest of the
microscope. It serves as a convenient “handle” when carrying a
microscope.

8. Body Tube – This hollow cylinder has an ocular lens at the

upper end and a revolving nosepiece, to which the objective
lenses are attached, at the lower end.
9. Draw Tube – Situated on top of the body tube,
it increases magnification.

10. Coarse Adjustment Knob – A wheel

operating by rack and pinion gear, the coarse
adjustment changes the elevation of the tube
rapidly for preliminary focusing. The coarse
focusing knob has a larger diameter and is used
to bring objects into rough focus.
11. Fine Adjustment Knob– This smaller wheel
moves the tube up and down gradually and
permits accurate and final focusing. The
fine focusing knob has a smaller diameter and is
used to bring objects that are in rough
focus into sharp focus.

12. Dust Shield - A circular disc above the

revolving nose piece, it protects the
objectives from dust particles.
13. Revolving Nose Piece – A rounded structure
carrying three objectives, the lenses may be swung
into their respective positions. The nosepiece may be
rotated to move an objective into viewing position.

14. Mechanical Stage. The mechanical stage is the

device on the stage surface that holds the slide
and enables precise movement of the slide. It is
operated by the mechanical stage control
knobs.
 II. OPTICAL PARTS
1. Ocular (Eye piece) – This cylindrical structure is
inserted into the upper end of the draw
tube.
2. Objectives – Cylindrical tubes attached to the
revolving nose piece for the initial
magnification of the image, they are
adjusted to be parfocal, that is, when any
objective is swung into its position, it is so
nearly focused as to require only the
use of the fine adjustment.
A. Low Power Objective (LPO, 10:1) – The LPO
views the specimen in a larger field. The shortest
objective is the low power objective, and it has a
magnification of 10X.

B. High Power Objective (HPO, 45:1) – A greater

magnification is obtained with this objective
where detailed structures may be observed. The
intermediate length objective is the high-dry
objective, with a magnification of 45X.
C. Oil Immersion Objective (OIO, 100:1) –
Greater magnification than the HPO is further
obtained by using this objective. A drop of oil
is placed on the slide for better refraction. The
longest objective is the oil immersion
objective, which has a magnification of 100X.
The magnifications of the oil immersion and
high-dry objectives may vary slightly in different
models of microscopes.
III. ILLUMINATING PARTS
1. Mirror – This is located below the stage to reflect light
rays through the object. One side of the mirror is
plane while the other is concave. The former is used
with natural light while the latter is employed when
artificial light is available. The mirror is supported by a
mirror arm swinging like a pendulum and having at its
end a mirror fork rotating vertically. This facilitates
movement in any desired position or angle.
2. Substage – Consisting of two parts, it is used
primarily for focusing light rays upon the
specimen.

A. Condenser – A combination of lenses beneath

the stage, the condenser makes it
possible for a maximum amount of light to be
directed on the object. It concentrates the light
on the microscope slide.
B. Iris diaphragm – This is made up of a
number of thin pieces of metal operated by a
lever for regulating the amount of light
necessary to obtain a clear view of the
specimen. It regulates the amount of light
reaches the slide.
Magnification
The microscope lens system magnifies
specimen so that very small structures can be
distinguished. The total magnification is
determined by the power of the ocular and
objective being used. It is calculated by
multiplying the power of the ocular by the power
of the objective. For example, a 10x ocular and a
45x objective yield a total magnification of 450x.
Resolution

Magnification without resolution is of little value.

Resolution is the ability to distinguish tiny
adjacent objects as two distinct objects.
Resolving power is a function of the wavelength
of light and the design of the microscope lenses.
The shortest wavelengths (blue) of visible light
provide maximum resolution. This is why
microscope have a blue light filter over the lamp.
Use of the oil immersion objective is required
for maximum resolution, and on the best light
microscopes it will enable the distinction of
microscope objects that are 0.2.µm apart. If
they are closer together, they will be seen as
one object because of a fusion of the images.
Focusing
A microscope is focused by changing the
distance between the object on the
microscope and the objective lens. This
change is accomplished by using the coarse
and fine focusing knobs. The focusing
knobs raise and lower either the stage or
the body tube depending on the type of
microscope. Both types of focusing
procedures are described below. Determine
As a general rule, you should always start focusing
with the low power (10X) objective. The coarse
focusing knob is used only with the low power
objective. With the high-power and oil immersion
objectives use only the fine focusing knob.
Switching Objectives

Modern microscope are usually parcentric and

parfocal. This means that when an object is centered
in the field and in sharp focus with one objective, it will
be centered and in focus when another objective is
rotated into the viewing position. However, it may be
necessary to make slight adjustments to re-center the
object with the mechanical stage or bring it into sharper
focus with the fine focusing knob.
 Start your observations with the low power
objective, even though you may want to observe
the object with the high-power or oil immersion
objectives. Once the object is center and in focus
with the low power objective, it is easy to switch
to the high-power objective simply by rotating the
nosepiece. Note that the working distance, the
distance between the objective and the slide,
decreases as the power of the objective increases.
 The amount of light entering the objective
decreases as the power of the objective
increases. Thus, you will need to increase the
voltage or open the diaphragm a bit to provide a
light intensity that yields the sharpest image with
the high-dry objective.
 The easiest and safest way to bring the oil immersion
objective into position is to progress from low power to high-
dry to oil immersion. However, you may switch directly from
the low power objective to the oil immersion objective.
Before rotating the oil immersion objective into the viewing
position, place a drop of immersion oil on the slide. Also,
open the diaphragm to its maximum aperture to increase the
amount of light. A slight adjustment of the fine focusing
knob is all that will be required to bring the object into focus.
Lens Care
Develop the habit of cleaning the lenses with lens
paper before using the microscope. Use only lens paper
for cleaning the lenses. If liquid gets on the objectives
or stage of the microscope, wipe it off immediately with
lens paper. If simply wiping the lenses with lens paper
doesn’t get them clean, it may be necessary to clean
them with lens paper moistened with green soap or
xylene. Use such a procedure only as directed by your
instructor. Never try to disassemble any part of the
microscope.
The best way to determine if the ocular is clean is
to rotate it between your thumb and forefinger
while looking through the microscope. A rotating
pattern is evidence of a dirty lens. If cleaning the
top lens fails to remove the debris, inform your
instructor. If the ocular is removed to clean the
lower lens, it is imperative that a piece of lens
paper be placed over the open end of the body
tube.
An image that appears cloudy or blurred indicates
a dirty objective. If cleaning with lens paper
moistened with water fails to clear the image,
consult your instructor about cleaning the lens
with xylene.
Routinely wipe off the upper surface of the
condenser lens with lens paper to remove any
accumulated dust.