A microscope is an essential instrument in any scientific laboratory.

It is used for observation of structural details of cells, tissue or organ in sections.

A microscope is an optical apparatus consisting of one or more lenses in order to enlarge and/or
magnify minute objects.
A modern compound microscope has following structural components.
Non-Optical Components:
1. Base (foot):
It is horseshoe or U–shaped metallic structure that holds the whole microscope.
2. Pillar:
It is a short straight part that joins to base as well as arm.
3. Arm (Limb):
It is a metallic curved holder that attaches with the arm by inclination joint.
It supports body tube and stage.
4. Inclination Joint:
It is used for angling the microscope if essential for observation in sitting position.
5. Stage:
It is a platform with a central hole fitted to the lower part of the arm. Microscopic slides held on the
stage by either simple side clips or by a mechanical stage clip.
6. Body tube:
It is meant for holding objective and ocular lenses at its both sides.
The end containing ocular lens is called head whereas the end holding 3 to 4 objective lens is called
nose piece.
The body tube has an internal space for the path of light rays which form the enlarged microscopic
particles.
7. Draw tube:
It is a narrow tube that fixed at the upper part of the body tube. It connects ocular lens or eyepiece.
8. Rack and pinion:
The microscope has a rack and pinion mechanism in body tube for moving the object under focus.
9. Adjustment screws:
There are two pairs of screws; they are large and small, larger for coarse adjustment and smaller
for fine adjustment. In fine adjustment the body tube or stages moves for extremely short distances.
Whereas in coarse adjustment is long distance from the object.
Optical Components:
10. Diaphragm:
It is fluttered just below the stage for regulating the amount of light falling on the object. Diaphragm
is of two types, disc and iris.
11. Condenser:
It is attached beneath the diaphragm. Condenser can be moved up and down to focus light on the
object.
12. Reflector (Mirror):
It is connected just above the base. Both its surface consists mirrors, a concave on one side and
plane on another side. The concave side is used in weak light and plane side in strong light.
Reflector directs the light on the object by the condenser and diaphragm system.
13. Objective Lenses:
They are fitted over the nose piece. Objective lenses are of two 10 three types – low power (10X or
5X), high power (45X) and oil immersion (100X). An objective lens is a compound lens. It produces
the real inverted image of the object inside the body tube.
14. Ocular Lens or Eyepiece:
An ocular or eyepiece lens is so called because it is normally the lens that is adjacent to the eye
when someone views through the device.
The eyepiece is of four types, they are 5X, 10X, 15X, and 20 X. The advanced microscope has two
eyepieces so that both the eyes can be used.

Working Principle of Compound Microscope:

The compound microscope is most commonly used in clinical, scientific and educational
laboratories.
The object or specimen, to be observed is normally fixed on a transparent glass slide and placed on
the specimen stage between the objective lens and condenser lens.
A ray of visible light from the base is focused by a condenser lens onto the specimen. The objective
lens accumulates the light transmitted through the specimen and produces a magnified image of the
specimen called primary image inside the body tube. This image is further magnified by the
eyepiece or ocular lens.
When higher magnification is required, the nosepiece is wheeled after low power brings on
producing the objective of the higher power in line with the illuminated part of the slide.
The objective lens should not touch the slip or coverslip.
Only fine adjustment can help for proper focusing. More light may be required. After observation
under high power, the nosepiece is rotated to bring back the slide under low power.
Oil immersion is a technique used to increase the resolving power of a microscope
When high magnification is required (e.g. for observing bacterial cell) oil immersion objective lens
(usually 100X) is employed.
Placing a drop of oil with the same refractive index as glass between the objective lens and cover
slip removes two refractive surfaces, so that magnifications can be attained while still preserving
good resolution.
phase microscope (phase-contrast microscope) a microscope that alters the phase relationships of
the light passing through and that passing around the object, the contrast permitting visualization
of the object without the necessity for staining or other special preparation.
Dark field microscope one so constructed that illumination is from the side of the field so that
details appear light against a dark background.

Electron microscope
A microscope with high resolution and magnification, using electron beams in place of light and
using electron lenses is called electron microscope.
The first electron microscope was developed by Ernst Ruska and Max Knoll of Germany in 1931.
EM can magnify an object up to 1000000X (one million times).
Light microscope can achieve a maximum resolution of about 0.2 µm or 200 nm, whereas EM can
achieve a resolution of 0.10 nm which is 2000 times better than the light microscope. Resolution is
the ability of a lens to distinguish between closely positioned small objects.
Principle of Electron Microscope:
Electrons are subatomic particles, which orbit encompassing the atomic nucleus. When atoms of a
metal are excited by heat energy, electron particles fly off from the atom. In the electron
microscope, tungsten is heated by using a high voltage current, electrons form a continuous stream,
which is used like a light beam.
The lenses used in EM are magnetic coils, which is capable of focusing the electron beam on the
specimen and illuminating it. Greater the flow of the current, greater will be a strength of the
magnetic field.
Transmission Electron Microscope (Tem):
Transmission electron microscopy (TEM) is a microscopy technique in which a beam of electrons is
transmitted through an ultra-thin specimen, interacting with the specimen as it passes through it.

It consists of a system of electromagnetic lenses fixed in a column.

Components of Electron Microscope:
It consists of the following main components:

1. Electron gun
2. Electromagnetic lenses—three sets.
3. Image viewing and recording system.
Electron gun is a heated tungsten filament, which produces electrons.
Condenser lens focuses the electron beam on the specimen.
A second condenser lens forms the electrons into a thin tight beam.
To move electrons down the column, an accelerating voltage is applied between tungsten filament
and anode.
EMs use accelerating voltages between 100 kV-1000 kV.
The specimen to be examined must be remarkably thin, at least 200 times thinner than those used
in the optical microscope. Ultra-thin sections of 20-100 nm are cut. The specimen holder is an
extremely thin nitrocellulose paper supported by a metal grid.
The electronic beam passes through the specimen and electrons are scattered depending upon the
thickness of the specimen. The denser regions in the specimen scatter more electrons; in contrast,
transparent regions are brighter.
The electron beam coming out of the specimen passes down the second of magnetic coils called
objective lens, which has high power and forms the intermediate magnified image. Finally, the third
set of magnetic lenses called ocular lenses produces the final magnified image.
Each of these lenses acts as image magnifier with an incredible level of resolution. This image is
projected on a fluorescent screen. Below the fluorescent screen is a camera for recording the image.
As the EM works in the vacuum, the specimen should be completely dry. Air molecules present in
the column of EM scatter the electrons causing flicker in the electron beam.

Image Viewing and Recording Techniques:

The highly magnified image is formed below the projector lens on a fluorescent screen. Below this
screen, a camera or a film or light sensitive sensor such as charge-coupled device (CCD) camera are
placed. The image can be displayed on computer or monitor. For direct viewing monocular or
binocular viewing, lenses are used.

Scanning Electron Microscope:

A scanning electron microscope (SEM) is a type of electron microscope that generates images of a
sample by scanning it with a focused beam of electrons.
Therefore SEM is used to study the surfaces of the microbes in excellent detail.
Secondary electrons hit a scintillator which transmits light which is transformed into electrical
current and amplified. The signal is sent to a cathode ray tube and forms image in the computer
monitor.
Electron beam passes through a pair of scanning coils or deflector plates in the electron column of
the SEM in the final lens which deflects the beam on the surface of the specimen. The electrons
interact with atoms on the surface of the specimen producing signals that contain information
about the properties of the specimen.
The signals produced in a SEM include secondary electrons, backscattered electrons (BSE). X-rays,
transmitted electrons and light. The signals scan the surface of the specimen from side to side, in
lines from top to bottom. This is known as raster scanning. They generate high-resolution images of
the specimen surface and produce information about the composition of the specimen.
Use of Electron Microscope:
The invention of EM has come as a boon for biological sciences and industry.
Immense magnification, a high resolution has opened new prospects for investigation in cellular
and molecular biology.
Study of microorganisms like the virus, bacteria and other pathogens have made the treatment of
diseases very effective. Fields of medicine, pathology, human anatomy have gained immensely from
electron microscope studies. Health field has benefited tremendously. Nanotechnology studies are
the result of electron microscope studies.
The science of microbiology owes its development to the electron microscope. It also helps in
tumour identification, biopsy, a study of cells, and the variety of molecules.

Difference # Light Microscope:

1. Visible light is used in this microscope.
2. The source of illumination is located at the bottom.
3. For magnification in the light microscope, the lens system consists of glass lenses.
4. The lenses are ocular, objective and condenser.
5. The image is either seen with the eye or recorded on a photographic film.
Difference # Electron Microscope:
1. Electrons are used in this microscope.
2. The source of illumination is situated at the top in this microscope.
3. The lens system consists of electromagnetic coils in this microscope.
4. This microscope has projector coils, an objective and a condenser.
5. The image in an electron microscope is either recorded on a fluorescent scent screen or recorded
on a photographic film.