AMITY INSTITUTE OF BIOTECHNOLOGY

AMITY UNIVERSITY MUMBAI

CELL BIOLOGY and GENETICS LAB

B. Tech Biotechnology, Semester 1
Experiment Number: 2

Title: Microscopy

Aim:
To study different types of Microscopes.

Introduction: A Microscope is an instrument used to see objects that are too stroll to
be seen by naked eye. It is derived from the Greek word "micro"
meaning small and "scope" meaning to look/see/aim.

Microscope is the science of investigating small objects and

structures. The most common microscope is optical microscope which
uses light to pass through a sample to produce an image. The image
obtained is known as Micrograph.
Chromosomes at either end of the cell begin to cluster
together which facilitates the formation of new nuclear membrane. This is
also when cytokinesis occurs leading to two separate cells.

Types of Microscopy

1. Magnifying lens/ simple microscope

It is a microscope with one lens.

2. Compound Microscope

It is a microscope with two lens. Two parameters especially

important in compound microscope are:

a) Magnification - It is the measure of how much larger a

microscope causes an object to appear.

b) Resolution - Resolution of microscope or lens is the smallest

distance by which two points can be separated and still be
distinguished as separate objects.
The smaller the value, the higher the resolving power of the microscope
and better the clarity.

Both magnification and resolution are important to get a clear picture of

something very tiny.
Structural Components of Compound Microscope

The three basic structural components of compound microscope are

Head, Base and Arm.

a) Head/Body houses the optical parts in the upper part of the

microscope.

b) Base of the microscope supports the microscope and houses the

illuminator.

c) Arm connects to the base and supports the microscope head. It is

also used to carry the microscope. When carrying a compound
microscope always take care to lift it by both the arm and base
simultaneously.

Optical components

There are two optical components in a compound microscope: Eye

piece lens and Objective lens

a) Eye Piece/Ocular Lens

It is what you look through at the top of the microscope. Typical

standard eye piece has magnifying power of 10X. Optional eye
piece of various powers are available typically from 5X to 30X.

b) Eye Piece Tube

It holds the eyepiece in place above the objective lens.

c) Objective Lens

They are primary optical lens on a microscope. They range from

4X to 100X and typically include three, four or five lens on most
microscope. It can be forward or rear facing.
 d) Nose Piece

It houses the objective lens. The objectives are exposed and are
mounted on a rotating turret so that different objectives can be
conveniently selected. Standard objectives include 4X, 10X, 45X
and 100X, although different power objectives are available.

e) Coarse and Fine Focus Knobs

They are used to focus the microscope. They are co-axial knobs
i.e. to say they are built on the same axis with the fine focus knob
on the outside. Coaxial focus knobs are more convenient since the
viewers does not have to grab a different knob.

f) Stage

It is where the specimen to be viewed is placed. A mechanical

state is used when working at higher magnification where delicate
movements of the specimen slide are required.

g) Stage Clip

They are used when there is no mechanical stage.

h) Aperture

It is the hole in the stage through which light reaches the stage.

i) Illuminator

It is a light source for the microscope typically located in the base

of the microscope. Most light microscope use low voltage halogen
bulbs.

j) Condenser

It is used to collect and focus the light from the illuminator onto
the specimen.

k) Iris Diaphragm

It controls the amount of light reaching the specimen.

 l) Condenser Focus Knob

It moves the condenser up or down to control the lighting focus of

the specimen.

3. Fluorescence microscope

It is used to image the samples that fluoresce.

4. Confocal Microscopy

It uses a laser to excite a thin layer of sample.

5. Electron microscopy

a) Transmission Electron Microscopy

It works on a similar principle to an optical microscope, but uses

electrons in place of light and electromagnet in place of glass lens.

b) Scanning Electron Microscope

It produces an image of the sample by scanning the surface with

the focused beam of electrons.

c) Atomic Force Microscopy

It provides picture of atoms on / in surfaces. It can provide higher

resolution than scanning electron microscopy.

d) Scanning Tunneling Microscopy

It is a non-optical microscope that works by scanning an electrical

probe tip over the surface of a sample at constant spacing. This
allows for a 3D picture of the surface to be created.

e) X-ray Microscopy

It uses electromagnetic radiation in the soft X Ray band to produce

magnified image of objects.
Requirements:

Procedure: 1.
2.
3.
4.

Observations:

Conclusion/
Inference/ Different types of microscopes were studied.
Approach: