MICROSCOPY

UNIT-I
 PURPOSE AND TYPES
• Microscopy is a
technique use for making
very tiny things to visible
to the naked eyes and
the instrument used to
make things visible to
the unaided or naked eye
is known as Microscope.
• The microscope is the
instrument most
frequently
characteristics of
microbiology laboratory.
• The magnification
provides by the
microscope enables us
to see microorganisms
and their structures
otherwise invisible to
the naked eye.
 PRINCIPLES OF MICROSCOPE

1.Magnification
2.Resolving power
3.Numerical aperture

• Magnification: It is the ability of lenses to enlarge an object visually.

If the magnifying power of lens is 10X it means that the given lens can
enlarge the object up to 10 times. In compound microscope, the
magnification is the product of magnifying power of both the lenses.
The magnifying power of lens depends on focal length. Lower the
focal length, higher is the magnifying power of lens.
• Resolution: The resolving power is the ability to distinguish two closely
placed points. The resolution power of lens allows us to observe the details of
an object. The resolution of microscope can find out by using Abbe’s
equation. Where,
d – distance between two closely distant points
λ – wavelength of light
n sin θ – numerical aperture
The microscope with higher magnification has small d value. λ is the
wavelength of light, shorter is the wavelength; higher is the resolution. The
wavelength of visible light is from 300 to 700 nm. The best resolution for
light microscope is obtained in the range of 450 to 500 nm. ‘n’ is the
refractive index of medium. Refractive index is the ability of the medium to
bend the light. The angle of cone of light is affected by the refractive index of
medium. The refractive index of air is 1. ‘θ’ is the half of the angle of the
cone of light that enters the microscope. The value of ‘Sin θ’ cannot be more
than 1 because angle of entering cone of light cannot be more than 90° and
value of sin 90 is 1.
TYPES OF MICROSCOPES
• Antony Van Leeuwenhoek invented the
first simple microscope in 1683. In
simple microscope, single lens is used
for magnification whereas in compound
microscope, multiple lenses are used.
Monocular microscope has one eyepiece
and binocular has two eyepieces. As
research has advanced, we now have
different types of microscopes that are
commercially available.
 PARTS OF MICROSCOPE
1. Base: The bottom part is called as base. It gives
support to the body. The illuminating source of
microscope is placed at the base of microscope
2. Arm: It connects the ocular part with the base.
The arm is used to hold and carry the
microscope. The coarse and fine focusing knobs
are placed on the arm.
3. Eyepiece: It is present at the top of microscope.
It is the part through which we observe the
sample/object. The magnifying power of the
eyepiece lens is generally 10X.
4. Eyepiece tube or body tube: The function of
eyepiece tube is to hold the eyepiece and hence
it is names as eyepiece tube.
5. Nosepiece: This part connects the eyepiece tube to objective lenses. The flexibility of nosepiece allows
switching the objective lenses.
6. Objective lens: In compound microscope, in general 3 objective lenses are placed. Their magnifying power
is 10X, 40X and 100X respectively. The total magnification of respective lenses would be 10X x 10X = 100X,
40X x 10X = 400X and 100X x 10X = 1000X (the magnifying power of eyepiece lens is 10X)
7. Focusing mechanism (Adjacent knobs): The coarse and fine adjustment knobs are used for right placing
of sample and for focusing it.
8. Stage: It is place where sample or object is kept for viewing. For accurate placement, the stage is provided
with clips that hold the slide firmly. The adjustment knobs are used to move the clips as per requirement.
9. Aperture: It is present on the stage that allows the entry of light from illuminator to fall on sample or object.
10. Source of Illumination: In non-electric microscope, the sunlight is used as source of illumination and
hence mirror is placed to focus the sunlight. In electric microscope, lamp is placed of specific wavelength.
11. Condenser: It is placed at the bottom of the stage. Its function is to focus the light on sample form
illuminator. The lens quality of condenser equally affects the superiority of image.
12. Diaphragm: It is generally associated with condenser. And it is also placed at the bottom of the stage,
beside condenser. The diaphragm and condenser together produce hollow cone of light that strikes the sample
and illuminate it.