Compound microscope

Komar University of Sciences and Technology

Objectives
1. Learn the parts of a compound light microscope and their
functions.

2. Learn how to calculate the magnification of a compound light

microscope.

3. Learn the proper use of the low and high-power objective lenses.

4. Learn the proper use of the coarse and fine adjustments for
focusing to find images.
 background
• Microscope is a technology of making
very small things visible to naked eye.

• The word microscope is derived from the

Greek mikros (small) and skopeo (look at).

• In 1672, Leeuwenhoek developed a first

simple microscope with a magnification of
200X – 300X.

• He is called as Father of microscopy

Types of microscopes
• There are many types of microscopes, and they may be grouped in
different ways.

• One way is to describe the way the instruments interact with a sample
to create images, either by sending a beam of light or electrons to the
sample.
Types of microscope
❑ Light Microscopes

1. Simple or dissecting microscope (stereo or stereoscopic microscope: low

magnification, 3D image).
2. Compound microscope:
✓ Bright-field microscope (is the simplest microscopy techniques; simplicity makes it a popular
technique)
✓ Fluorescence microscope (uses UV)
✓ Inverted microscope (useful for observing living cells or organisms at the bottom of a large
container)
✓ Confocal microscope (for increasing optical resolution and contrast of a micrograph )
✓ Phase-contrast microscope (reveals many cellular structures that are invisible with a bright-field
microscope)
❑ Electron Microscope (magnification power
of 100,000,000 X)

1. Scanning Electron Microscope (SEM)

2. Transmission Electron Microscope (TEM)
Compound microscope and its parts:
• Light will be used as source of illumination

• The microscope consists of a stand (base + neck), on which is

mounted the stage (for holding microscope slides) and lenses.

• Two sets of lenses; the ocular lens (or eye piece) which magnifies an
object 10 times its normal size, and the objective lenses located on a
revolving nosepiece.
Calculation of Total Magnification
• Each objective lens has a different magnification of power written on
it (such as 4, 10, 40, or 100).

• For total magnification multiply the ocular power times (10X) by

the objective lens.

• For example, if you have a 10X ocular and a 10X objective, the total
magnification is: 10X × 10X = 100X.
• Positioning the specimen requires that you turn the mechanical stage
controls.

• One control moves the specimen in the x-direction, and the other
moves the specimen in the y-direction.

• Focusing on the specimen is achieved by knobs that move the stage

up and down, so that it is closer or farther from the objective.

• There are two knobs, an outer coarse focus and an inner fine focus.

• The substage condenser directs light through the slide into the
objective
Using a compound microscope
1. Open the clip on the stage and put the slide on the stage.
2. Use the knobs to centre the specimens over the condenser
lens.
3. Start with low power objective lens (4X lens). Rotate the
coarse adjustment knob to find a clear image.
4. Move to the next objective lenses (10X and 40X) and
refocus using fine adjustment knob.
5. NEVER use the coarse adjustment for lenses higher than
4X
Using immersion oil
1. The 100X objective lens (oil immersion objective lens) needs to be used with oil

2. Focus on the specimen with the “high dry” lens (high power objective lens
40X).

3. Rotate the objective lens somewhere between the 40X and 100X lenses
4. Place a drop of immersion oil on the cover slip, and very carefully swing the oil
immersion lens into place.

5. Adjust the fine focus until you get a crisp and clear image.

6. after using100X objective lens, carefully wipe the oil from the lens
Note:

• Make sure oil NEVER touches the 40X objective lens.

• Oil will ruin the 40X objective lens!
• After using a microscope return the objective to a “start” position
(4X).
• Never leave the 100X objective in place when returning the
microscope to the cabinet.
Cleaning the lens of a compound microscope

• 1. Dampen a piece of lens paper with 1 to 2 drops of lens cleaning

solution.
• 2. Hold the dampened lens paper against the lens for 5 to 10 seconds.
Thanks for your attention