INTRODUCTION

MICROSCOPE: A microscope is an instrument used to see the objects that are too small for the

naked eye. Such type of object

are called microscopic object.
And the science of investigating
of small objects using such type of instrument is called microscopy.

There are many types of microscopes. The most common (and the first to be invented) is the
optical microscope, which use light to image the sample. It consists of lens or a combination of
lens which produced a magnified image of the object under study. It is an indispensable tool for
cytologists. Other major types of microscope are electron microscope, ultra microscope and
various types of scanning probe microscope.

Between 1590 and middle of 17th century several improvements were made in terms of
mmagnification and the refinement of lenses. ‘Anton van Leeuwenhoek’ (1674), the father of
biology, was the first to use the microscope for biological study. His microscope was consisted
of a single lens with high power of magnification. The compound microscope was constructed
by ‘Robert Hooke’ (1665) and is the forerunner of the present day microscope. Many of
Leeuwenhoek’s reports were confirmed by Robert hook. As refinements and improvements in
the construction of microscope continued, more and more detailed description of the object
become available become available. In 1830 achromatic lenses were made. Good quality
microscopes with oil immersion lenses become available only in 1870. However, refinements
continued to be made, e.g. addition of a binocular eyepiece, built in illumination and
photomicrography facility.

POLARIZATION MICROSCOPE
The polarization microscope utilize plane polarized light in order to study the presence of
preferentially-oriented constituents, the direction of their orientation, their shape, regularity of
internal construction and refractive index in the cell and tissues.
The image formation in the polarizing microscope is based on the unique ability of polarized
light to interact with ordered molecules in a direction-sensitive manner. Perturbations to waves
of polarized light from aligned molecules in an object result in face retardation, which in term
allow interference dependent changes in amplitude in the image plane. Thus, image formation
is based not only on principle of diffraction and interference, but also on the existence of
ordered molecular arrangements. In the polarizing microscope, such structures generally
appear bright against a dark background. The polarizing microscope is also a remarkable analytical
instrument, capable of providing quantitative measurements of differences in optical path length
(retardation), which in turn can be used to calculate refractive index differences and the thicknesses of
ordered specimens. Geologists use these parameters together with a reference chart to
determine the identities of unknown crystalline minerals. These capabilities distinguish
polarization microscopy from other forms of light microscopy and account for its popularity in
biology, chemistry, geology, and material science.

PRINCIPLE OF POLARIZATION MICROSCOPE

The polarization microscope is based on the behavior of some cell components when observed
under polarized light. The cell components may be either isotropic or anisotropic.

ISOTROPIC SUBSTANCE: Isotropic substances are those through which polarized light is
propagated with the same velocity irrespective of the impinging direction. The isotropic
property of a substance is due to its similar index of refraction in all directions.

ANISOTROPIC SUBSTANCE: Anisotropic substances are those through which polarized light is
propagated with varied velocity. This anisotropic property is due to two different indices of
refraction of the material. The two indices of refraction correspond with two different velocity
of transmission is turned as birefringence. Many transparent crystals and minerals such as
quartz, calcite, tourmaline and others are optically anisotropic and exhibit double refraction
phenomena. Therefore these substances are called doubly refracting or birefringent.
Birefringent materials split an incident ray into two components that traverse different
paths through the crystal and emerge as two separate rays. This occurs because
atoms in crystals are ordered in a precise geometric arrangement causing direction dependent
differences in the refractive index. When a ray of light is incident on a birefringent crystal, it
usually becomes split into two rays that follow separate paths. One ray, the ordinary ray or O
ray, observes the laws of normal refraction, while the other ray, the extraordinary ray or E ray,
travels along a different path. Thus, for every ray entering the crystal there is a pair of O and E
rays that emerges, each of which is linearly polarized
Figure 1
Double refraction in a calcite crystal. A letter viewed through a plane surface of the crystal
appears double, the two images corresponding to the ordinary and extraordinary rays. As the
crystal is rotated, the E ray rotates around the O ray. The O ray obeys the normal laws of
refraction and does not rotate.

the ability of molecularly ordered objects to split an incident ray of light into two components,
the O and E rays, but the two terms refer to different aspects of the same process. Double
refraction refers to the visible phenomenon: the splitting of a single incident ray into two
resultant rays as exemplified by a crystal of calcite. Birefringence refers to the cause of
the splitting: the existence of direction-dependent variation in the refractive index in a
molecularly ordered material. Birefringence B also refers to a measurable quantity, the
difference in the refractive index (ne- no) experienced by the O and E rays during transit
through an ordered object such that
B = (ne - no).

Depending on the values of ne and no, the sign of birefringence may be positive or
negative, and specimens are therefore said to be either positively or negatively birefringent.
Birefringence is related to the relative retardation.