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..
