THE MICROSCOPE
• Illuminator and Stage
Identify the basic parts of the microscope
• Describe each part according to their functions
• Explain the working principle of the microscope
• Manipulate virtually a compound microscope
• Enumerate the proper handling of microscope
COMPOUND MICROSCOPE DEFINITION
• The term microscope can be split into two separate words,
‘micro’ and ‘scope’, where the term
‘micro’ means small or tiny, and ‘scope’ means to view or to
observe. Therefore, a microscope
can be understood as an instrument to observe tiny elements.
• The optical microscope often referred to as the light
microscope, is a type of microscope that
uses visible light and a system of lenses to magnify images of
small subjects.
• There are two basic types of optical microscopes-simple and
compound.
• The term “compound” in compound microscopes refers to the
microscope having more than one
lens.
• Devised with a system of combination of lenses, a compound
microscope consists of two optical
parts, namely the objective lens and the ocular lens
ARTS OF THE COMPOUND MICROSCOPE
• Eyepiece and Body Tube.
The eyepiece is the lens through which the viewer looks to see
the specimen.
It usually contains a 10X or 15X power lens.
The body tube connects the eyepiece to the objective lenses.
• Objectives and Stage Clips
Objective Lenses are one of the most important parts of a
Compound Microscope.
They are the closest to the specimen.
A standard Microscope has three to four Objective Lenses which
range from 4X to 100X.
Stage Clips are metal clips that held the slide in a place.
• Arm and Base
The Arm connects the Body Tube to the base of the Microscope.
The Base supports the Microscope and its where Illuminator.
The illuminator is the light source for a microscope.
A compound light microscope mostly uses a low voltage bulb as
an illuminator.
The stage is the flat platform where the slide is placed.
• Nosepiece and Aperture
Nosepiece is a rotating turret that holds the objective lenses.
The viewer spins the nosepiece to select different objective
lenses.
The aperture is the middle of the stage that allows light from the
illuminator to reach the specimen.
• Condenser, Iris diaphragm, and Diaphragm
A condenser gathers and focuses light from the illuminator onto
the specimen being viewed.
Iris diaphragm adjusts the amount of light that reaches the
specimen.
The diaphragm is a five holed disk placed under the stage.
Each hole is of a different diameter. By turning it, you can vary
the amount of light passing through the
stage opening.
WORKING PRINCIPLES OF THE COMPOUND MICROSCOPE
• Compound microscopes have a combination of lenses that
enhances both magnifying powers as
well as the resolving power.
• The specimen or object, to be examined is usually mounted on
a transparent glass slide and
positioned on the specimen stage between the condenser lens
and objective lens.
• A beam of visible light from the base is focused by a condenser
lens onto the specimen.
• The objective lens picks up the light transmitted by the
specimen and creates a magnified image
of the specimen called the primary image inside the body tube.
This image is again magnified by
the ocular lens or eyepiece.
• When higher magnification is required, the nose piece is
rotated after low power focusing to
bring the objective of a higher power (generally 45X) in line with
the illuminated part of the
slide.
• Occasionally very high magnification it required (e.g. for
observing bacterial cell). In that case, an
oil immersion objective lens (usually 100X) is employed.
• The common light microscope is also called a bright-field
microscope because the image is
produced amidst a brightly illuminated field. The image appears
darker because the specimen or
object is denser and somewhat opaque than the surroundings.
Part of the light passing through
or object is absorbed.
MAGNIFICATION OF THE COMPOUC MICROSCOPE
• In order to ascertain the total magnification when viewing an
image with a compound light
microscope, take the power of the objective lens which is at 4x,
10x or 40x and multiply it by the
power of the eyepiece which is typically 10x.100x-OIO
• Therefore, a 10x eyepiece used with a 40X objective lens will
produce a magnification of 400X.
The naked eye can now view the specimen at magnification 400
times greater and so
microscopic details are revealed.
• Alternatively, the magnification of the compound microscope is
given by:
m = D/ fo * L/fe
where, D = Least distance of distinct vision (25 cm)
L = Length of the microscope tube
fo = Focal length of the objective lens
fe = FHOW TO FOCUS • SOME IMPORTANT DO’S:
. Always start with the scanning or low power objective. Turn the a. ALWAYS use the scanning or low power objective first. Get
nosepiece to rotate this objective into the slide in focus with this objective,
position and listen for the “click”. If you do not hear the “click”, then switch to a higher power. Do this is for every slide you view,
the objective is not all the way in not just the first slide of the lab period.
position. b. Only use lens paper on the lenses and slides.
b. Use the coarse adjustment knob to bring the scanning or low c. Treat your microscope gently.
power objective as close to the slide as d. Ask questions when it’s not clear how to do something.
possible. Watch the objective from the side when you are • SOME IMPORTANT DON’TS:
lowering the objective to prevent breaking a. NEVER use the coarse adjustment knob when using high
the slide or objective. power. Since this objective is so long
and the coarse adjustment moves it so fast, there is great
c. Look through the eyepiece and turn the coarse adjustment danger of breaking the slide or lens.
knob to slowly RAISE the objective AWAY PROPER TECHNIQUE: On high power, use only the fine
from the slide. STOP when the image comes into focus. adjustment knob to focus.
d. Refine your focus by using the fine adjustment knob. Note that b. NEVER focus downward with the coarse adjustment while
when you manipulate the fine looking through the eyepiece. PROPER
adjustment you are actually focusing at different levels with the TECHNIQUE: You should lower the low power or scanning
specimen. This gives you depth objective first while viewing from the side (not
perception of the material being viewed. the eyepiece) and then focus upward while looking through the
e. Move the stage so that the part of the slide you want to view is eyepiece.
in the center of the field. c. DON’T try to find an object under high power that you cannot
f. Experiment with the amount of light passing through the locate under low power.
specimen by moving the lever on the iris PROPER TECHNIQUE: First locate the object under low power
diaphragm. You will need to adjust the iris diaphragm for each and focus. Next move the stage so that
slide and each magnification. the object is in the center of the field of view and then switch to a higher
PROPER HANDLING OF MICROSCOPE power.
a. These microscopes are delicate, expensive instruments and
have to be treated with care.
b. Always carry the microscope with two hands; one hand
supporting the base and the other hand on
the arm.
c. Only use lens paper to clean the lenses. The glass used in
microscope lens is softer than ordinary glass
and it easily scratches. Do NOT use facial tissues, paper towels
or any cloth to clean the lens. If you get
water on the lens, wipe it off with lens paper. Your fingers are
oily; NEVER touch the lenses.
d. When you finish using the microscope, remove the slide from
the stage and turn the nosepiece so that
the scanning objective is in position. Turn off the light switch and
pull out the plug. Wrap the cord
around the base of the scope.
e. Put the microscope away at its correct location. Match the
microscope number (on the arm) to its
corresponding number within the cabinet.

HOW TO CHANGE MAGNIFICATION?

a. Always start with the scanning or low power objective in
position. Focus with this objective and
center your slide as described above.
b. Rotate the nosepiece to bring a higher power objective into
position. Listen for the “click”.
c. These microscopes are parfocal which means that if an object
was in focus and centered with low
power, it should also be in focus or near focus with high power.
Use only the fine adjustment knob with
the high-power objective. NEVER focus downward with the
coarse adjustment knob when using high
power; a sure way to crack a slide.
d. Adjust the iris diaphragm so that the amount of light passing
through your specimen gives the
maximum contrast. Usually the brightest light (maximum amount
of light) provides little contrast. You
need to make this adjustment for every slide.

local length of the eye-piece lens

RULES IN MICROSCOPE USE