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THE MICROSCOPE
PARTS OF THE
MICROSCOPE _________
LIGHT SOURCE
It serves as a support for microscopes.
Illumination for the modern microscope is
Microscope illuminators are also carried by it.
by a light source located in the base. Light square or horseshoe-shaped base
source is equipped with a ____________ to
regulate the intensity of the light. Filters may ARM
also be placed to vary the illumination and
wavelengths of the emitted light. It joins the base to the head and the eyepiece
tube to the base. It supports the microscope's
CONDENSER head and is also used for carrying the
instrument. Entire upper assembly of the
______________on the specimen and
microscope is held in an upright position
controls/concentrates the light for uniform
illumination. BODY TUBE
_____________ body tube holds the ocular in place:
Although called a "tube." it may be more like a
Adjust the light intensity, controls the
box in some models.
amount of light and the angle of light rays
that pass to the specimen and lens, which ____________________
affects resolution, contrast, depth of the field
of image. A turretlike circular mechanism rotates so
that different lenses can be selected. Always
OBJECTIVE rotate the nosepiece by holding the outside of
Perform the ___________ magnification of the revolving disk.
the object on the mechanical stage.
OBJECTIVES
OCULAR (EYEPIECE) 4× (_____________) is used for the initial
location of the specimen.
FURTHER magnify the object that has
10x (____________) may also be used for the
been enhanced by the objects for viewing, initial location of the specimen. It is also used
gives additional magnification to the object. for observing specimens that don't need greater
Set of lenses closest to your eye. magnification.
40x (_____________) is used for specimens
COURSE ADJUSTMENT KNOB requiring greater magnification. This objective
Moves the mechanical stage noticeably up is called dry because it does not require the use
and down until the object comes into view of oil, as other high-power objectives do.
for initial focusing. 100x (____________) is used for magnification
of extremely small specimens p, such as
FINE ADJUSTMENT KNOB bacterial cells. It must be immersed in oil, so it
adjust to ______ the image. is called the high oil objective.
STAGE
The specimen is usually mounted on a
glass or plastic microscope slide that rests on
the stage, a platform just below the objective.
The stage has a hole so that light can
pass through the specimen from below. If the
stage has an adjustable bracket that
moves the slide around mechanically by
rotating positioning knobs, the stage is
called a mechanical stage. If not, the slide is
held by stage clips and must be moved
by hand.
_____________________
Below the stage is a high intensity lamp.
Light rays from the lamp travel through a
hole in the stage, through the specimen
mounted on a slide, then through the
objective and ocular, to the eye.
TOTAL MAGNIFICATION
Total magnification is determined by multiplying the power of the ocular
by the power of the objective in use. Thus, when using a 10X ocular and
the low-power objective, total magnification is 100x.
TYPES OF
MICROSCOPY
Light Microscopy
• A microscope is an instrument that
magnifies an image and allows
visualization of greater detail than is
possible with the unaided eye
• Two types:
________________________
________________________
SIMPLE MICROSCOPE
a scientific instrument used to
magnify microscopic objects so that they
are visible to the unaided eye. It is a
double convex lens with a short focal
length that is mounted in a lens holder.
Note***
The first simple microscope was created
by Dutch scientist Antonie Van
Leeuwenhoek to examine the tiny
microorganisms in freshwater.
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COMPOUND MICROSCOPE ______________ MICROSCOPE
high magnification microscope that uses 2 is used to display autofluorescent
lenses to compound (multiply) the level of molecules such as vitamin A and some
magnification. The first lens is referred to as neurotransmitters. Widespread application
the objective lens and typically has a 4x, is in the detection of antigens or
10x, 40x or 100 magnification ability). The antibodies in immunocytochemical
second lens is known as the eyepiece lens. staining procedures
PHASE CONTRAST MICROSCOPE CONFOCAL SCANNING
MICROSCOPE
enables examination of unstained cells
and tissues and is especially useful for living allows visualization of a biologic
cells. specimen in three dimensions. It combines
components of a light optical microscope
Two modifications of the phase contrast with a scanning system to dissect a
microscope are: specimen optically
Interference microscope, which also
ULTRAVIOLET MICROSCOPE
allows quantification of tissue mass
uses quartz lenses with an ultraviolet
Differential interference microscope
light source.
(using Nomarski optics), which is especially
useful for assessing surface properties of The image depends on the absorption
cells and other biologic objects. of UV light by molecules in the specimen.
may achieve a resolution of 0.1 m and
DARK-FIELD MICROSCOPE resembles the workings of a
useful in examining autoradiographs, in spectrophotometer; results are usually
which the developed silver grains appear recorded photographically
white in a dark background.
POLARIZING MICROSCOPE
Clinically it is useful in examining urine
for crystals and in demonstrating specific a simple modification of the light
bacteria microscope in which a polarizing filter
particularly Treponema pallidum. (the polarizer) is located between the light
source and the specimen, and a second
polarizer (the analyzer) is located
between the objective lens and the viewer
It uses the fact that highly ordered
molecules or arrays of molecules can
rotate the angle of the plane of polarized
light.
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ELECTRON MICROSCOPY
Two kinds of EMs can provide
morphologic and analytic data on cells and
tissues: bacteria particularly Treponema
pallidum.
transmission electron microscope
uses the interaction of a beam of
electrons with a specimen to produce an
image
scanning electron microscope
the electron beam does not pass through
the specimen but is scanned across its surface
CARE OF MICROSCOPE
o Always use two hands to carry the scope, one on the arm, one on the base.
o Never touch the lenses with your fingers.
o Do not force knobs
o Keep objects clear of desk and cords
o Lens paper is the only tissue that can be used when cleaning the eyepiece.
o Be careful when plugging the power cord into the outlet.
o Always plug it into a receptacle. at your station, taking care not to sting it over a chair, across
an aisle, or in any other dangerous position
o Make sure the stage and objective are at their farthest distance apart and
that the lowest-power objective is in position. Start each new observation at low power.
o At the end of the lab, the stage should be placed all the way down, an objective lens placed on
scanning power (red) when putting away.
o Make sure the stage is in its lowest position and wrap up the cord neatly wound the base and
place the cover over the top of the microscope when finished.
o Always grab the appropriate microscope for the station you are at and put it away in the
appropriate spot in the cabinet.
o Be careful when plugging the power cord into the outlet.
o Always plug it into a receptacle. at your station, taking care not to
sting it over a chair, across an aisle, or in any other dangerous position
o Make sure the stage and objective are at their farthest distance apart and that the lowest-power
objective is in position. Start each new observation at low power.
o When rotating the objective lens, do not rotate from HPO to OIO; it should be LPO to
scanning going to OIO. To prevent the HPO lens from contacting the oil.
USING THE MICROSCOPE
1. Obtain a microscope slide
2. Carefully place your slide on the stage and secure it with stage clips or the brackets
of the mechanical stage. Move the slide so that the specimen is centered in the
stage's hole.
3. While looking from the side, use the coarse-focus knob to move the objective (still
on low power) as close as possible to the slide.
4. Look through the ocular and use the coarse-focus knob to slowly move the
objective and slide apart.
5. When the image becomes clear, switch to the fine-focus knob to make the image
even sharper.
6. Adjust the light intensity by using the lamp controller or diaphragm until the detail
of the image is at its clearest.
7. Notice that the sample appears upside down and backwards in your field of view.
This inversion is caused by the way the lenses in the scope bend light to magnify
the image. You will notice the same effect if you hold an ordinary magnifying glass
at arm's length.
8. Center the part of the specimen that you wish to see more clearly and switch to a
higher-power objective:
Notice that as you move the slide to the left, it appears to move to the right.
This reversal is caused by the lenses in the scope. Most scopes are parfocal, which
means that the image remains focused when you change objectives. However. some
minor adjustment with the fine focus is usually necessary.
9. Practice focusing at different depths by using a prepared slide
10. When you remove the slide, make sure that the low-power objective is in the
viewing position and that the objective and stage are as far apart as possible.
11. When you have finished for the day. return the scope to its original configuration.
The low-power objective should be in position. Make sure the slide has been removed.
PREPARED BY: LOVELY DIANE V. DAMOT, RMT
AND NIKOLAI L. MARTIN, RMT
REFERENCES:
Wise, E.(2022). Microscopy. In E. Wise, Laboratory Manual Essentials
of Seeley’s Anatomy and Physiology. (10th ed). New York: McGraw
Hill Education.
VanPutte, C. L. (2019). Seeley’s Anatomy and Physiology.
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