Serial No. : - 01 Date: - 14.01.

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STUDY OF COMPOUND MICROSOPE

 Aim of the experiment: - To study the parts of compound microscope.

 Principle: - The typical component of a compound microscope is capable of increasing the ability to
see the details by thousand times. So that an object as small as 1 micrometer can be seen this
microscope. The electron microscope cans extent the ability up to 0.1 micrometer.
A focus of light passing through the spectrum and that having a high refractive index of color stain. A
shadow which is magnified in 2 nanometer as it passes the observer’s eye. A compound microscope is
called so because in contrast to a simple magnifying convex lens.

It has two such lenses: - I) Objective lens II) Eye piece

It magnifies the image of an objective that is not visible to the naked eye to an extent where it can be
visible clearly.

 Parts of microscope:

1) Eye piece: - Microscope usually has 5x, 10x & 15x magnification.
 
2) Both tubes: - Body tube contains mirror and prism which directs the image to the eye piece.
 
3) Nose Piece: - It holds objective lens and rotates.

4) Objective Lens: - Usually microscope has 3 or 4 objective lenses i.e. 10x, 45x, 100x lens, sometimes
some manufacturers also provide 4x magnification lens.

a) LOW POWER (LP):- [10x; N.A= 0.25; Focal Length=l6mm]

The lower power objective in common is used to magnify 10 times. It is used for
initial focusing and viewing a large area of specimen slide. The N.A of this lens is always greater than
that of the condenser in most microscopes. In order to achieve the focus, therefore the N.A has to get to
be closely matched by reducing the light reaching the specimen under study. This is achieved by
lowering the condenser and partially closely to the iris diaphragm.

b) HIGH POWER (HP):- [45x; N.A = 0.65; Focal Length = 4 mm]

The lens magnifies the image 45 times because of this higher magnification. This is
used for more detail study of material before switching to oil immersion lens. The N.A of high power
lens is almost equal to slightly less than that of the commonly used condenser. Therefore the later has to
be slightly raised and the iris diaphragm opens to get more light and maximum clarity in focal length.

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 c) OIL IMMERSION OBJECTIVE (OI):- [100x; N.A=1.38; Focal length = 2mm]
The oil immersion objective lens magnifies the image slightly. The lens almost
touches the slide. It has to be immersed in a special media mostly into wood oil. A drop of which is first
placed on the slide. The oil is used to increase the N.A and thus the resolving power of the objective.
Since the N.A of oil immersion objective is always greater than that of the condenser, Later has to be
raised to the highest position and the iris diaphragm is fully open. As the lens gives total magnification
of 1500 times. It is required for the detailed study of blood cells and tissues.

5) Stage: - Movable plot on which the studies are mounted.

6) Condenser: - A sub stage lens which focus the light on the specimen.

7) Iris Diaphragm: - A diaphragm is located below stage to control light and direct effect of the focus
on the image.

8) Focusing knob: - Use to focus the object up and down movement.

9) Light Source: - It is either located or built inside the microscope or may be external power supply.
The plane mirror is used for the OI objective whereas the concave mirror used for LP & HP objective.

Compound Microscope
Definitions
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 1. Resolution: The limit of useful magnification of a microscope is set by its resolving power, that is, its
ability to revel closely adjacent structural details as separate & distinct. Resolution therefore describes
how small individual objects can lie close to each other and still be recognizable. The light microscope
can separate dots that are 0.25 µm apart.
The resolving power is expressed quantitatively as microscope’s limit of resolution (LR), that is, the
minimum distance between two visible bodies at which they are seen as separate and not in contact with
each other. The LR is determined according to the following formula:
LR =0.61 X W/ NA [Where, W= wave length of light rays
NA = numerical aperture of the objective in use]

2. Working Distance: Working distance is the distance between the objective & the objective slide. The
working distance increase with decrease magnification.
 
3. Numerical Aperture (NA): the numerical aperture of a lens is the ratio of the diameter of the lens to
its focal length. Any particular lens has an constant NA and this value is dependent on the radium of the
lens & its focal length. As the NA increase, the resolution decrease. Greater the NA .the greater the
resolving power. The NA is also describe as an index of the light gathering power of a lens, that is the
amount of light entering the objective.

 PROCEDURE FOR THE USE OF MICROSCOPE:-

 
1) Turn the light source and raise the body tube 7-8 cm above the stage.

2) Switch to 10x objective lens.

3) Adjust the focus to raise specific lower lenses.

4) Keep the slide in proper position.

5) Look at the ocular lens, one lens is fixed and the other lens has a focusing ring like a binocular.
 
6) Bring lens close to the slide as possible then look at any fixed ocular lens then back up until specimen
come in focus then adjust the focus.
 
7) Looking through the ocular lens, adjust its focus. Look in with both eyes to see single light field and
make mirror adjust to the focus first.
 
8) Reset and adjust focus and readjust diaphragm as needed.

The best illumination can attend when we use this rule to adjust the microscope:

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Objective Condenser Iris
Position Diaphragm
   
Low Power (10x) Low Partly Open
 
High Power (45x) Midway Half Open
     
Oil-Immersion (100x) High Fully Open

DIAGRAM OF COMPOUND MICROSCOPE