MICROSCOPE

Microscope is an instrument that produces enlarged images of small objects, allowing the observer an
exceedingly close view of minute structures at a scale convenient for examination and analysis. Although optical
microscopes are the subject of this article, an image may also be enlarged by many other wave forms,
including acoustic, X-ray, or electron beam, and be received by direct or digital imaging or by a combination of these
methods. The microscope may provide a dynamic image (as with conventional optical instruments) or one that is static
(as with conventional scanning electron microscopes).
The magnifying power of a microscope is an expression of the number of times the object being examined
appears to be enlarged and is a dimensionless ratio. It is usually expressed in the form 10 (for an image magnified 10-
fold), sometimes wrongly spoken as ten eksas though the were an algebraic symbolrather than the correct form,
ten times. The resolution of a microscope is a measure of the smallest detail of the object that can be observed.
Resolution is expressed in linear units, usually micrometers (m).
The most familiar type of microscope is the optical, or light, microscope, in which glass lenses are used to form the
image. Optical microscopes can be simple, consisting of a single lens, or compound, consisting of several optical
components in line. The hand magnifying glass can magnify about 3 to 20. Single-lensed simple microscopes can
magnify up to 300and are capable of revealing bacteriawhile compound microscopes can magnify up to 2,000. A
simple microscope can resolve below 1 micrometer (m; one millionth of a meter); a compound microscope can resolve
down to about 0.2 m.
Other types of microscopes use the wave nature of various physical processes. The most important is
the electron microscope, which uses a beam of electrons in its image formation. The transmission electron
microscope (TEM) has magnifying powers of more than 1,000,000. TEMs form images of thin specimens, typically
sections, in a near vacuum. A scanning electron microscope (SEM), which creates a reflected image of relief in a
contoured specimen, usually has a lower resolution than a TEM but can show solid surfaces in a way that the
conventional electron microscope cannot. There are also microscopes that use lasers, sound, or X-rays. The scanning
tunneling microscope (STM), which can create images of atoms, and the environmental scanning electron
microscope (ESEM), which generates images using electrons of specimens in a gaseous environment, use other physical
effects that further extend the types of objects that can be examined.

TYPES OF MICROSCOPES
There are two different types of microscopes:
light and electron. Light microscopes have glass lenses
which magnify objects, and use light to illuminate the
objects being examined. Electron microscopes use
beams of electrons to examine incredibly small objects
(like the components of an individual cell) that have
been specially prepared. Light Microscope has 2
different types the Compound and the Dissecting
microscope.

The Compound Microscope
Compound microscopes are used to examine
objects in two dimensions. Very small organisms or
cross-sections of organisms are placed on clear glass
slides; these objects are viewed as light passes through
them. The parts of the compound microscope are
reviewed below.

The Dissecting Microscope
Dissecting microscopes are used to observe
material that is either too thick or too large to be
viewed with the compound light microscope. With
these microscopes, you see the surface of things that
reflect the light. While the magnification and depth of
field are smaller in the dissecting scope, the field of
view is much larger. As its name implies, the dissecting
scope is often used to look at plants as you dissect
them, since it allows for manipulation of material. Since
most of the parts of the dissecting microscope are the
same as the compound microscope, they will not be
reviewed here.

Electron Microscopy
In these microscopes a beam of electrons (in
place of light) and circular magnets (in place of glass
lenses) permit the resolution of structures in much finer
detail than in an optical microscope. There are two
electron microscopes. The first is a "traditional"
transmission electron microscope (TEM) in which an
electron beam passes through the specimen. The
second is the scanning electron microscope (SEM) in
which a beam of electrons scans the surface of an
opaque object and produces an image of that surface.
The images are viewed on a cathode tube, or in pictures
taken with the microscope. Many of the photographs of
cell structure used in your text were taken with an
electron microscope. FIU hosts the Florida Center for
Analytical Electron Microscopy, which has an SEM.


RULES IN USING THE MICROSCOPE
1. Always carry the microscope in a straight upright
position with one hand around the arm and the other
hand under the base. The eyepieces are not attached
and will fall out if the microscope is carried at an angle
or upside down.
2. Check out the microscope to make sure all the lenses
are clean and the mechanical parts are in working
order. Report any malfunction to the instructor so that
it may be remedied.
3. Keep the microscope clean. When anything is spilled
or otherwise gets on the microscope, clean it up
immediately.
4. When using the microscope start with the low power
lens and work up to the desired magnification. These
microscopes are par focal, which means that all powers
should be in focus when the turret is rotated.
5. Never move the stage upwards with the coarse
adjustment while viewing through the eyepieces. Get
the lens close to the slide while viewing from the side to
make sure that they never touch. Then move the stage
downward with the coarse adjustment while viewing
through the lens. This will prevent the possibility of
ramming the lens into the slide, thereby ruining a slide
you have just made and, quite possibly, damaging the
lens.
6. Moist, living or preserved materials must be observed
through a coverslip. This protects the lens as well as
tends to make the object under view optically flat. Be
sure to maintain a safe distance between the coverslip
and the objective lenses.
7. Clean the lenses with lens paper only. DO NOT CLEAN
THE LENSES WITH HANDKERCHIEFS, FACIAL TISSUES,
PAPER TOWELS, ETC.--they will scratch the lenses. If
your lenses are very dirty, obtain some lens cleaning
solvent from the instructor.
8. If you cannot obtain clear focus or good lighting, or if
your microscope seems not to be working properly,
IMMEDIATELY CALL YOUR INSTRUCTOR. He/she can
either assist you or see that the microscope is repaired.
9. Return your scope to the cabinet with light cord
wrapped around its base and with the lowest power
objective lens in position.

Parts of the compound microscope
A compound microscope is composed of many
important, functional components. There are at least
two lenses the ocular lens and objective lens. The
ocular lens is in the eyepiece; it is usually 10x power.
The objective lenses are near the stage on the revolving
nosepiece. There are usually multiple objective lenses
that are used to vary the magnification; on many
microscopes the objectives are 4x, 10x and 40x
magnification. The condenser and iris diaphragm are
important devices used to adjust how light hits the
sample, thus influencing the visualization of the sample.


Using a compound microscope
To use a microscope, you must know how to
focus on the sample, adjust the condenser and
diaphragm to optimize visualization of the sample. For
binocular scopes, you must also set the
appropriate inter pupillary distance and adjust
the diopter ring to focus both eyes on the sample.
The image you see through the microscope is
inverted, which should be kept in mind when moving
the stage and sample. The field of view and depth of
field decrease as the magnification increases; by
knowing the diameter (mm) of the field of view for each
objective lens, the size of objects can be measured.



Stereoscopic (dissecting) microscopes
A stereoscopic or dissecting microscope is a
relatively low magnification (often 2x-30x) microscope
that is good for viewing large objects. As its name
implies, it is better than a compound microscope for
dissecting many materials (e.g. small animals, plants,
organs) because it produces a 3-dimensional image.
Although its magnification is generally less than
that of a compound microscope, it has a couple of
advantages over compound scopes. First, it has a large
depth of field, so you can observe thick objects with
most parts in focus at the same time. Second, you can
illuminate your sample using incident light as well as
transmitted light, so that thick objects which will not
transmit much light can be illuminated adequately to
view.
Dissecting microscopes have an objective lens
which often allows a continuous range of magnification
(from 2-30x), controlled by a magnification knob. There
is a focus knob, and a source of transmitted light
(adjustable); many scopes also contain a source of
incident light. Another difference from a compound
microscope is that the stage is much farther from the
objective lens, allowing large objects to be placed on
the stage.


Electron Microscopes
An electron microscope visualizes objects with
electrons as the source of illumination, allowing much
greater magnification and resolution than a compound
microscope. Transmission electron microscopes
transmit electrons through a thin slice of sample.
Scanning electron microscopes use incident electrons
on whole samples to get a 3-dimensional image.
Similar to compound microscopes, electron
microscopes have an energy source (electrons instead
of light), an objective lens, and a condenser lens. Images
are viewed through an eyepiece, or projected onto a
screen or photographic plate.

The following images show: a Golgi apparatus at
unknown magnification,

the head region of a deer tick magnified 500x,

Spirogyra (Protista-green algae) magnified 500x,

and a diatom magnified 15,000x.

These images show a mosquito at increasing
magnification.