MICROSCOPIC ANATOMY
The Compound Microscope
Generally, cells of plants and animals are too small to be seen with the naked eye, thus
necessitating the use of a special instrument. Such an instrument is a microscope, which may be
of two general types:
1. the light microscope, which makes use of visible light.
2. the electron microscope, which makes use of a beam of electrons rather than visible
light
The compound microscope is a n example of a light microscope that makes use of two sets of
lenses:
1. The primary magnifier, the objectives, the lenses of which are
immediately next to the specimen being studied.
2. The secondary magnifier, 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. microscopic 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
b. capacity of the eyepiece and the objective being used
Example: high power objective - 40x
Ocular - 10x
Total power - 40 x 10
= 400
This means that the image is 40 times as the object under study.
4. units of measure
a. micrometer ( replaced the micron) – unit of measure of alight
microscope which is equivalent to 10 –3
b. manometer – 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.
NOTE: When the 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 of cone of light rays capable of entering the
front lens of the objective from a 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
a. 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
100x 1.25
NOTE: The higher the magnification of the objective, the larger is the numerical aperture
(NA)
10. resolving power (RP)
a. the ability of the lens to clearly separate or distinguish two points or
two lines individually in the image.
b. Determined by:
- wavelength of the light source
NOTE: The shorter the wavelength, the greater the resolving power numerical aperture
(NA)
NOTE: The greater the numerical aperture of the lens, the greater is resolving power
Mechanical Parts
1. Base – a horseshoe or V-shaped structure that 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
- serves as the handle for carrying the microscope as well as
providing a support for the body tube
4. Inclination joint
– movable hinge that facilitates tilting of the microscope by
holding the arm with your right hand and the base with
your left hand
5. Body tube – a hollow cylinder in front of the upper part of the arm and
serves as the optical housing for the lenses
- serves as a passageway of light from the objective to the
eyepiece as well as an attachment of the draw tube onto
the microscope
6. Draw tube – the smaller cylinder attached to the upper part of the body
tube and serves to hold the ocular the eyepiece
7. Resolving nosepiece
– a rotating head attached to the base of the body
tube and holds the different objectives and facilities shifting
of such objectives
8. Dust shield - a fixed head attached to the base of the body tube
but situated above the revolving nosepiece and
protects the objective lenses by keeping off dust
9. Stage - a platform, which may be circular or square, were the
slide with the specimen is placed; usually equipped with a
pair of clips that hold the slide in place
10. Substage condenser with iris diaphragm
- a structure found below the stage, holding the Abbe
condenser (see illuminating part below) above and the iris
diaphragm below
11. Adjustment knobs – consist of two pairs of knobs
a. coarse adjustment knob or screw – an upper larger pair is used for initial
focusing by raising or lowering the body tube, with the scanner or the low power
objective in position
b. fine adjustment knob or screw – the lower, smaller pair 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 while focusing with the high-power
objective or any high power objective. Use only the fine adjustment screw.
The coarse adjustment screw can be used when the low power or scanner objectives are
in place.
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 light into the specimen
- the concave mirror is used with low intensity light which may
be either natural or artificial the plane mirror is best used
when the source of light is very bright
2. condenser (Abbe condenser)
- the substage optical element which may be fixed or provided
with adjustment for lowering and raising
- functions to condense and concentrate reflected light from
the mirror through the specimen at a precise angle to
illuminate the microscopic field.
Magnifying Parts
1. eyepiece or ocular
a. the detachable cylinder equipped with lenses and located
above the draw tube
b. 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 (LPO)
- the shortest cylinder or tube with large lens opening,
large lenses and lower magnification
- used to observe the general view of the specimen as
well as to locate various parts of the specimen under
study
b. high power objective (HPO)
- the longer cylinder or tube with smaller lens opening,
smaller lenses and higher magnification
- used to observe the details of parts of the specimen
but very much enlarge
c. oil immersion objective
- the longest cylinder or tube with a very small lens
opening, very small lenses and very high magnification
used to observe fine details of specific parts and at a
very much higher magnification
d. scanner
- if there are four objectives present, then this would be
the shortest cylinder with the biggest opening and the
lowest lens opening and the lowest magnification
- best used when your specimen is big, and you would like
to view the entire specimen within the microscopic field
