THE COMPOUND MICROSCOPE

Generally, cells of plants and animals are too small to be seen with the naked eye, thus, necessitating the
use of special instrument. Such instrument is a microscope, which may be of two general types: a) the light
microscope which makes use of visible light; b) the electron microscope, which makes use of a beam of electrons
rather than visible light.
The compound microscope is an example of a light microscope that makes use of two sets of lenses:
a) a primary magnifiers, the objectives, the lenses of which are immediately next to the specimen being studied;
and b) the secondary magnifiers, the ocular or eyepiece, the lenses of which are immediately next to the eye.
Important terms in microscopy:
1. Virtual image or floating image – image seen in the microscope field.
2. Microscope field – circular area seen through the ocular where the image of the specimen is observed.
3. Magnification – ratio of the apparent size of the object, as seen through the microscope, and the actual size of
the object.
a. total magnification or total power – product of the magnifying capacity of the eyepiece and the
objective being used.
Ex. high power objective 40x
Ocular 10x
Total power 40 x 10 = 400
This means that the image is 400 times as big as the object under study.
4. Units of measurement
a. micrometer (m) – unit of measurement of a light microscope which is equivalent to 10 -6.
b. nanometer (m) – unit of measure of an electron microscope that is equivalent to 10 -9.
5. Focusing – the adjustment of the spatial relationship between the objective lens and the specimen. When a
clear image of the specimen is seen through the ocular lens, the specimen is said to be “in focus”.
6. Focal length of focus – the distance from the optical center of the lens to the focal point.
7. Working distance – the distance between the front (bottom lens) of the objective lens system and the top of
the cover glass.
Note: Lenses of high magnification and short focal lengths have a short working distance.
8. Angular aperture – the angle or cone of light rays, capable of entering the front lens of the objectives from the
point in the object.
Note: More light rays from the specimen can be taken in by the lens if the angular aperture of the
objective is increased, thus increasing also the resolving power.
9. Numerical Aperture (NA) of an objective – the measurement of the angle of the maximum cone of light that
may enter the lens, or simply a measurement of the ability of the condenser and objective lens to gather light.
Objective Numerical Aperture
4x 0.10
10x 0.25
20x 0.40
40x 0.65

Note: The higher magnification of the objective, the larger is the numerical aperture (NA).
10. Resolving power (RA) – the ability of the lens to clearly separate or distinguish two points or two lines
individually in the image.
11. Depth of focus – represents the thickness of the specimen that may be seen in focus at one time.
- decreases with an increase in the numerical aperture and magnification.
12. Parfocal – term applied to the objective lens when practically no change in focus has to when one power is
substituted for another.
Note: Only a slight turn of the fine adjustment is needed when a change is made from low power to a high
power objective.
13. Refraction – bending of light away from the objective lens when light passes from the glass of the microscope
slide to the air.
Note: Refraction is lessened by using oil (oil immersion objective).
14. Cover glass – thin glass (0.17 to 0.18 mm thick) cut in circles, rectangles or squares for covering the specimen.

PARTS OF A COMPOUND MICROSCOPE

I. MECHANICAL PARTS:
1. Base – horshoe or V-shaped structure which supports the entire microscope.
2. Pillar – vertical post extending from the base to the arm.
3. Arm – curved basic frame of the microscope to which the base, body and stage are attached.
- serve as the handle for carrying the microscope as well as providing support for the body tube.

4. Inclination joint – a movable hinge that facilitates tilting of the microscope by holding the arm with your right
hand and the bases with your left hand.
5. Body tube – a hollow cylinder in front of the upper part of the arm and the serves as the optical housing for
the lenses.
6. Draw tube – the smaller cylinder attached to the upper part of the body tube and serves to hold the ocular
eyepiece.
7. Revolving nosepiece – a rotating head attached to the bases of the body tube.
8. Dust shield – a fixed head attached to the base of the body tube.
9. Stage – a platform, which may be circular or square , where the slide with the specimen is placed.
- usually equipped with a pair of clips that hold the slide in place.
- Some microscopes are provided with a mechanical stage adjustment knobs for accurate
movement of the slide.
- Provided with central opening which allows light to pass from the mirror to the object being
studied.
10. Substage condenser with iris diaphragm – a structure found below the stage holding the abbe condenser
above and the iris diaphragm below. The iris diaphragm is provided with a projecting handle or lever which
when manipulated can regulate the amount of light passing through the lenses into the observer’s eye.
11. Adjustment knobs – consist of two pairs of knobs:

a. an upper pair, the coarse adjustment knob or screw which is used for initial focusing by raising or
lowering the body tube with the scanner or the low power objective in position.
b. A lower small pair, the fine adjustment knob or screw which is used for final viewing and adjustment
at different depths, with the high power objective in position.
Note: Never use the coarse adjustment screw when focusing with the high power objective or any high
powered objective. Use only the fine adjustment screw. The coarse adjustment screw can be used when
the low power or scanner objective is in place.

II. ILLUMINATING PARTS:

1. Mirror – usually two-faced, of which one surface is plane or flat while the other side is concave (parabolic)
- held by a double-hinged mirror rack or fork which facilitates adjustments at any desired
angle since the mirror functions to reflect the light into the specimen.
- The concave mirror is used with low intensity light which may be either natural or
artificial, while when the source of light is very bright, the plane mirror is best used.
2. Condenser (abbe condenser) – the substage optical element which may be fixed or provided with adjustment
for lowering or raising.
- functions to condense and concentrate reflected light from the mirror through the
specimen at a precise angle to illuminate the microscopic field.

III. MAGNIFYING PARTS:

1. Eyepiece or ocular – the detachable cylinder equipped with lenses and located above the draw tube.
- usually provided with dark pointer used to point particular parts of the specimen to be
identified.
2. Objective – generally with two or three objectives.
a. Low power objective – the shortest cylinder or tube with a large lens opening, large lenses
and lower magnification.
b. High power objective – the longer cylinder or tube with smaller lens opening, smaller lenses
and higher magnification.
c. Oil immersion objective – the longest cylinder or tube with very small lenses and very high
magnification.
d. Scanner – if there are four objectives present, then this would be the shortest cylinder with
the biggest lenses and lowest magnification.

Care of the Microscope

Microscopes are expensive tools, hence, must be handled very carefully at all times. The following are
some of the precautions that you must be familiar with when using them.

a. Examine your microscope before using it for any defects, like misplaced or missing parts. If there is
anything wrong with it, call the attention of the instructor, otherwise you will be held responsible for
any defects.
b. Keep it clean before and after using with a piece of soft cloth (not your handkerchief).
c. Do not separate loose parts, like the ocular or objectives, as they may collect dust.
d. Do not touch the lenses with your fingers. Use lens paper only to clean them.
e. Always check the parts thoroughly after receiving it from the counter and before returning.
f. If anything is spilled on the microscope, clean it off immediately.
g. Always carry the microscope carefully with one hand under the base and the other hand around the
arm. Never carry the microscope hanging like a basket with only one hand.