Microscopy
Dr B. Mahalakshmi
Eyepiece
Body Tube
Revolving Nosepiece
Arm
Objective Lens
Stage
Stage Clips
Coarse Focus
Diaphragm
Condenser Fine Focus
Light Base
Ocular lens
Magnifies 10 times
where you look through
to see the image of your
specimen.
They are usually 5X or
10X power.
Our microscopes have an
ocular lens power of 10x.
supports the tube
arm and connects it to
the base
The flat platform
where you place
your slides
stage
moves stage
(or body tube) up
and down
coarse adjustment
knob
Small, round knob on the
side of the microscope
used to fine-tune the
focus of your specimen
after using the coarse
adjustment knob
fine adjustment knob
the bottom of the
microscope, used
for support
base
The part that
holds two or
more objective
lenses and can Revolving
be rotated to nosepiece
easily change
objectives
Adds to the magnification
Usually there will be
3 or 4 objective
lenses on a Objective
microscope. lens
4X, 10X, 40X and
100X powers.
Objective lens coupled with eyepiece
lens, we get total magnifications of
40X (4X times 10X),
100X ,
400X and
1000X.
The shortest lens is the lowest power,
the longest one is the lens with the
greatest power. Lenses are color coded.
Magnification
Scanning, Low , high dry and oil immersion .
Each objective – magnification will be written.
The ocular lens (eyepiece) has a magnification.
The total magnification is the product
(multiplication) of magnifying power of ocular
and objective lens
Oil immersion 100X 10X 1000X
Stage clips hold the slides in place.
If microscope has a mechanical stage,
We can move the slide around by turning two
knobs.
One moves it left and right,
the other moves it up and down.
stage clips
Condenser
The purpose of the condenser is to
ensure that the objective is able to
provide the maximum resolution. This is
a system of different lens elements
which is mounted beneath the stage of
the microscope
Condensers can be
moved up and down.
The normal
operating position is Condenser
up.
Iris diaphragm
Diaphragm is used to vary the intensity and
size of the cone of light that is projected
upward into the slide.
By adjusting the
aperture diaphragm, it
is also possible to
Control depth of field
and contrast Iris
diaphragm
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.
Condensor
Makes the specimen
easier to see
Tungsten lamp
sunlight
illumination
Focusing Specimens
1. Always start with the scanning
objective.
Odds are, you will be able to see
something on this setting. Use the Coarse
Knob to focus and then the fine
adjustment knob until clear, image may be
small at this magnification, but you won't
be able to find it on the higher powers
without this first step.
Do not use stage clips, try moving the
slide around until you find something.
2. Once you've focused on Scanning, switch to Low Power.
Use the Coarse Adjustment Knob to refocus. Then use the Fine
Adjustment Knob to make the image crystal clear. Again, if you
haven't focused on this level, you will not be able to move to the
next level.
3. Now switch to High Power. At this point, ONLY use the Fine
Adjustment Knob to focus specimens.
Recap
1. Scanning --> use coarse and fine knob
2. Low power --> use coarse and fine knob
3. High power --> use fine knob only
DO NOT SKIP STEPS!!!!
Partially rotate so that 40x and 100x
objectives straddle the specimen.
Immersion oil
A special oil used in microscopy with
typically the 100x objective lens (usually
at 1000x total power). A drop of
immersion oil is placed on the cover slip
and the objective is lowered until it just
touches the drop. Once brought into
focus, the oil acts as a bridge between
the glass slide and the glass in the lens.
This concentrates the light path and
increases the resolution of the image.
Place a small drop of oil on the slide
in the center of the lighted area.
(Take care not to dribble on the
stage.)
Put the small drop
of oil directly over
the area of the
specimen to be
Examined.
Rotate so that the 100x oil
immersion objective touches the
oil and clicks into place.
Focus only with fine focus.
Hopefully, the specimen will come
into focus easily. Do not change
focus dramatically.
Clean up!:
When you have finished for the day, wipe
the 100x oil immersion objective carefully
with lens paper to remove all oil.
Wipe oil from the slide thoroughly with a
wipe.
Cleanse stage should any oil have spilled
on it.
The Light Microscope
Guidelines for Use
• Always carry with 2 hands
• Only use lens paper for cleaning
• Do not force knobs
• Always store covered
Troubleshooting
Occasionally you may have trouble with working your microscope. Here are
some common problems and solutions.
1. Image is too dark!
Adjust the diaphragm, make sure your light is on.
2. There's a spot in my viewing field, even when I move the slide the spot stays in
the same place!
Your lens is dirty. Use lens paper, and only lens paper to carefully clean the
objective and ocular lens. The ocular lens can be removed to clean the inside.
The spot is probably a spec of dust.
3. I can't see anything under high power!
Remember the steps, if you can't focus under scanning and then low power, you
won't be able to focus anything under high power. Start at scanning and walk
through the steps again.
4. Only half of my viewing field is lit, it looks like there's a half-moon in there!
You probably don't have your objective fully clicked into place..