MICROSCOPY:

Compound Light Microscope

PARTS OF COMPOUND MICROSCOPE
 A
A. Eyepiece
B B. Body Tube
C. Coarse Adjustment

F D. Fine Adjustment
E
H E. Arm

F. Revolving Nosepiece

G J G. High Power Objective

H. Low Power Objective
I C
I. Stage Clips
K D J. Stage
L K. Diaphragm
M L. Light Source (Lamp)
M. Base
A Compound Microscope has 2 Lenses

A compound light microscope uses two lenses to make an image look

bigger. One is the eyepiece lens and the other is the objective lens.
Each lens bends the light making the image look larger.
THINK! - What is the magnification number on the eyepiece?
10x
THINK! - What is the magnification on each objective lens?
Scanner: 4x
Low Power: 10x
High Power: 40x
Oil Immersion: 100x
 Compound Light Microscopy

In a compound
microscope, the
image from the
objective lens is
magnified again by
the ocular lens
Total magnification =
objective lens 
ocular lens

Figure 3.1b
Calculating Total Magnification
To figure out the total magnification use this equation:
Total Magnification (x) = Eyepiece Mag. x Objective Mag.
THINK! - Use the equation above to calculate total magnification.
Total Magnification under Scanner=
10 x 4 = 40x
Total Magnification under Low Power =
10 x 10 = 100x
Total Magnification under High Power =
10 x 40 = 400x
Total Magnification under Oil Immersion =
10 x 100x = 1000x
1. Stage and Stage clips:
A fixed platform with an opening
in the center allows the passage
of light from an illuminating source
below to the lens system above
the stage
This platform provides a surface for
the placement of a slide with its
specimen over the central
opening
most microscopes have a
mechanical stage that can be
moved vertically or horizontally by
means of adjustment controls
2. Ocular Eyepiece
the lens at the top
that you look through
Has a magnification
of 10x
3. Body Tube
 Connects the
ocular eyepiece
to the revolving
nosepiece
4. Revolving nosepiece
This is the part that holds
three or more objective
lenses and can be rotated
to easily change power.
5. Objective lenses
Combined with ocular lens
to increase magnification
Scanner, LPO,HPO, OIO
6. Arm
Connects the eyepiece
to the objective lenses
The part where to hold
and carry the
microscope
7. Course Adjustment Knob
Brings the specimen into
general focus.
Used in scanner and LPO
8. Fine Adjustment Knob
Brings the specimen
into fine and tune focus
and increases the detail
of the specimen
Used in HPO and OIO
9. Illuminator/Stage Light
The light source of the
microscope
Older microscopes
used mirrors to reflect
light from an external
source up through the
bottom of the stage
9. Condenser
Gathers and focuses
light from the illuminator
onto the specimen
being viewed.
10. Iris diaphragm
Adjusts the amount of light
that reaches the condenser
and the specimen.
The function of the
transparency of the
specimen, the degree of
contrast you desire and the
particular objective lens in
use.
11. On/Off switch
 This switch on the base of the
microscope turns the illuminator
off and on.
General Procedures

• Plug your microscope in to the

extension cords above and in front of
each lab table.

• Store with cord wrapped around

microscope and scanner objective
clicked into place.

• Carry by the base and arm with both

hands.
Focusing Specimens

1. Always start with the scanner

objective.

Odds are, you will be able to see

something on this setting.

Use the Coarse Knob to focus, 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.
“Scan” for it.
2. Once you've focused on Scanning, switch to Low Power.
Use the Coarse Adjustment Knob to refocus. 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 knob
2. Low power --> use coarse knob
3. High power --> use fine knob
4. OIO  use fine knob
DO NOT SKIP
STEPS!!!!
●Your slide MUST be focused on low power before
attempting this step
●Click the nosepiece to the longest objective
●Do NOT use the Coarse Focusing Knob, this could
crack the slide or the lens
●Use the Fine Focus Knob to bring the slide
 MAGNIFICATION vs. RESOLUTION

Magnification is the ability to make small objects

seem larger, such as making a microscopic
organism visible.
Resolution is the ability to distinguish two objects
from each other.
Light microscopy has limits to both
its resolution and its magnification.
 Compound Light Microscopy

The refractive index is a measure of

the light-bending ability of a medium
The light may bend in air so much
that it misses the small high-
magnification lens
Immersion oil is used to keep light
from bending
Refraction in the Compound Microscope

Figure 3.3
 Cleanup
1. Store microscopes with the scanning
objective in place.

2. Unplug and wrap cords around the back

of the microscope. Cover microscopes.

3. Double check to make sure you didn't

leave a slide.

4. Place microscopes in their designated

location (if instructed by the teacher).
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.
Troubleshooting
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..