Introduction

to the
Microscope
Introduction:
A microscope is a tool that allows us to view things
that are too small to see with the naked eye. The
most common microscope is the compound light
microscope.
It uses a system of two or more lens to collect and
focus the transmitted visible light through the
specimen to the eye.
Light Microscope - uses compound (2) lenses to
magnify objects. The lenses bend or refract light to
make the object beneath them appear closer.
Anatomy of a compound light
microscope
Magnification

Your microscope has 3 magnifications (objective lenses):

o
o
o Scanning
Low
Each objective has its magnification labeled. In addition to this, the _______ ______ (eyepiece)
High
has a magnification of __X.
To calculate total magnification, ________ the ocular x objective.

Ocular Lens 10
multiply
 Total
Magnification
Total magnification is the ocular magnification
multiplied by the objective magnification as shown in
the following table.
Objective Magnification Ocular Lens Total Magnification

Scanning 4x 10x 40x

Low Power 10x 10x 100x
High Power 40x 10x 400x
Oil Immersion 100x 10x 1000x
A typical compound light microscope is able to
magnify from 40x to 1000x, increasing the ability to
see small details in the specimen. Most microscopes
will have three to four objectives with differing
magnification. They generally have a scanning, low,
high, and oil immersion objectives. Each objective wi
be marked with it magnification.
The ocular lens also magnifies at generally a 10x
magnification. The power of magnification will be
marked on the lens. It may be monocular or binocular
(one lens for one eye or two for both eyes). The
majority of our scopes are binocular.
 Set up and Use
1. Remove the dust cover. Carry your microscope
with two hands, one under the base and the
other gripping the arm or frame to the
appropriate bench.
2. Gently place the microscope on a flat, level
surface and plug the microscope into a power
outlet.

3. Turn on the light source The on button may

appear on the back or side of the microscope.
4. Adjust the brightness of the light source. A low
setting is generally best for scanning a slide. This is
done by one of two ways depending on the
microscope that you are using.
One way to adjust brightness is to adjust it using the
brightness adjustment located on the side of the
microscope.
This model generally also has an adjustable
condenser. It is generally marked for the individual
objectives. Set the condenser accordingly.
On older microscopes the brightness is adjusted by
turning the Illumination Ring on top of the light source.

By turning the ring you can decrease the amount of

light or increase the amount of light being shown
through the sample
5. Adjust the stage as low as possible with the
coarse focus knob. It is the outer or larger
knob.

6. Rotate the nose of the microscope so that

the lowest objective is in line with the light
source.
7. Place a slide on the stage holding it with the slide
clip. If a slide has a coverslip –it should always face
up.
8. Once your slide is correctly seated, look through
the ocular lens, adjust your brightness and using the
coarse adjustment knob (outer knob) bring the stage
up towards the objective, stopping when the slide
image comes into view. The slide should never be
pushed all the way up to the objective.
9. Once the image if seen use the fine focus
adjustment knob (inner knob) to sharpen in on the
details of the structure. After this point you will only
use the fine focus on the slide. All coarse adjustments
have been made.
With the scanning or lowest objective in use- you
may scan across the slide by moving the stage from
right to left and left to right. This is done using the
stage knobs found just below the stage of the
microscope.
As you scan across the slide you may need to use the
fine focus (inner knob) to sharpen the image. Once
you decide on an area of the slide you may want to
look at a higher magnification of the area.
This is done by rotating the nose of the microscope to
the next objective. On our microscopes that is a 10X
objective. Once the object has been rotated and
locked into place and you have made your light
adjustments (generally brighter); look through the
eyepiece and using only the fine adjustment knob
sharpen the image.
If the magnification still does not give the detail that
you need the nose of the microscope may be rotated
again to lock in the third objective which in our case
is a 40X. Again brightness may need adjusted and the
fine focus is used to focus and sharpen the image.
To view significantly small structures the oil
immersion objective (100x) may need to be used.
This is used by rotating the nose of the microscope to
between the third objective and the oil immersion
objective. At this point a small drop of oil is placed on
the slide and then the oil immersion objective is
rotated in passing through the oil on the slide.
If you get oil on the other objectives notify the laboratory
manager as soon as possible so that the objectives can be
removed and properly cleaned. If the oil is allowed to dry
on the objective it will not come off and all images viewed
thereafter will have a haze and not focus correctly.
10. When finished viewing your image- use the coarse
adjustment knob to lower (go away from) the objectives of
the microscope. Rotate the nose of the microscope to the
scanning or 4x objective. If oil has been used, use lens
tissue and cleaner to remove the oil from the slide,
objective, and stage.
11. Turn off the light and unplug the microscope. Cover the
microscope with a dust cover.
12. Return the microscope to the shelf.
13. If you encounter any issues tell or ask your lab
assistant, instructor or laboratory manager.
REMEMBER!

1. Always start with the Scanning Objective.

2. Place your specimen in the center of your field of view.
3. Use the Coarse Knob to focus.
4. Next, use the Fine knob until clear.
5. Once you've focused on Scanning, switch to Low Power.
6. Repeat steps 3-4.
7. Now, switch to High Power.
8. 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

What will happen if I use the course knob under high power?

The slide or lens could crack

Field of View

• Field of View: the area a person can _____

view through
a microscope.
circle
• It is represented by a _______.
increases field
• RELATIONSHIP: As magnification __________,
of view decreases
__________.
• As you switch from scanning (40X) to high power (100X),
the area you see through the microscope gets smaller.
• We can see better detail with higher powers of
magnification, but we cannot see as much of the image.
• The lens inverts the image of the object inside the
microscope. _________
Inverted means that the image
appears upside down and backward compared with
the actual object.
→

• This means that the slide must be moved in the opposite

direction that you want the image to move.
• Ex. If you move the slide to the right on stage, as you
look through the microscope the slide appears to
move ____.left
Making a Wet Mount Slide
1. Get a clean slide and coverslip from your teacher.
2. Place ONE drop of water in the middle of the
slide.
3. Place the specimen into the water.
4. Place the edge of the cover slip on one side of the
water drop at a 45° angle.
5. Slowly lower the cover slip on top of the drop.
THIS PREVENTS AIR BUBBLES.
Staining Specimens
Scientists stain specimens to see parts of the cell
more clearly.

1. Put a drop of stain (iodine) on a slide. Caution:

methylene blue will stain clothes and skin.
2. Place specimen in the center of drop of stain.
3. Place a coverslip onto the slide.
Drawing Specimens

1. Use pencil - you can erase and shade areas

2. All drawings should include clear and proper labels (and be

large enough to view details). Drawings should be labeled with the
specimen name and magnification.

3. Specimens should be drawn to scale - Ex. if your specimen takes

up the whole viewing field, make sure your drawing reflects that.
Practice

Drawing should have: Fewer cells at a larger size

Cleanup

1. Store microscopes with the scanning objective in place.

2. Wrap cords and cover microscopes.

*Double check to make sure you didn't leave a slide

3. Place microscopes in their designated location (probably a cabinet)

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. .