Bright Field Microscope

Submitted by :- Ritika Devi

Enrollment no. :- A0989220038

Course :- Basic microbial techniques

B.S.C Microbiology

Semester 1st

Submitted to :-

 Dr. Jagriti Tyagi ma'am

 Dr. Arti geol ma'am
 Dr. Menaka Devi Salma ma'am
Definition :-

Bright field Microscope is also known as the Compound Light Microscope. It is

an optical microscope that uses light rays to produce a dark image against a
bright background. It is the standard microscope that is used in Biology,
Cellular Biology, and Microbiological Laboratory studies.

This microscope is used to view fixed and live specimens, that have been
stained with basic stains which gives a contrast between the image and the
image background. It is specially designed with magnifying glasses known as
lenses that modify the specimen to produce an image seen through the
eyepiece.

Principle :-
For a specimen to be the focus and produce an image under the Brightfield
Microscope, the specimen must pass through a uniform beam of the
illuminating light. Through differential absorption and differential refraction,
the microscope will produce a contrasting image.

The specimens used are prepared initially by staining to introduce color for
easy contracting characterization. The colored specimens will have a
refractive index that will differentiate it from the surrounding, presenting a
combination of absorption and refractive contrast.

The functioning of the microscope is based on its ability to produce a high-

resolution image from an adequately provided light source, focused on the
image, producing a high-quality image.

The specimen which is placed on a microscopic slide is viewed under oil

immersion and covered with a coverslip.

Diagram :-
Parts and functions of bright field microscope :-

The brightfield microscope is made up of various parts, including

Eyepiece (Ocular lens) – it has two eyepiece lenses at the top of the
microscope which focuses the image from the objective lenses. this is where
you see the formed image from, with your eyes.
The objective lenses which are made up of six or more glass lenses, which
make a clear image clear from the specimen or the object that is being
focused.
Two focusing knobs i.e the fine adjustment knob and the coarse
adjustment knob, found on the microscopes’ arm, which can move the stage
or the nosepiece to focus on the image. Their function is to ensure the
production of a sharp image with clarity.
The stage is found just below the objectives and this is where the specimen
is placed, allowing movement of the specimen around for better viewing with
the flexible knobs and it is where the light is focused on.
The condenser: It is mounted below the stage which focuses a beam of
light onto the specimen. It can be fixed or movable, to adjust the quality of
light, but this entirely depends on the microscope.
The arm: This is a sturdy metallic backbone of the microscope, used to carry
and move the microscope from one place to another. They also hold the
microscope base which is the stand of the microscope. The arm and the base
hold all the microscopic parts.
It has a light illuminator or a mirror found at the base or on the
microscope’s nosepiece.
The nosepiece has about two to five objective lenses with different
magnifying power. It can move round to any position depending on the
objective lens to focus on the image.
An aperture diaphragm (contrast): It controls the diameter of the beam
of light that passes through the condenser. When the condenser is almost
closed, the light comes through to the center of the condenser creating high
contrast and when the condenser is widely open, the image is very bright
with very low contrast.

Magnification :-
The magnification of the image is determined by the magnification of the
objective against the magnification of the eyepiece lens. The objectives have
a magnification power of 40x-1000x depending on the type of brightfield
microscope while the eyepiece lens has a standard magnification power of
10x.
Therefore to calculate:
Total Magnification power = Magnification of the objective lens x
Magnification of the eyepiece

For example: if the magnification of the objective is 45x and that of the
eyepiece is 10x, the total magnification of the specimen will be 450x.
The magnification is standard, i.e not too high nor too low, and therefore
depending on the magnification power of the lenses, it will range between
40X and 100oX.
The objective lens enlarges the image which can be viewed, a characteristic
known as resolution. Resolution according to Prescott, is the ability of a lens
to separate or distinguish between small objects closely linked together.
Whereas the eyepiece magnifies the image at the end of the viewing, its
magnification range is lower than that of the objective lens at 8X-12X (10X
standard) and that of the objective lens at 40X-100X, magnification, and
resolution of the microscope is highly dependant on the objective lens.
Resolution of bright field microscope :-

'resolution' is used to describe the ability of a microscope to

distinguish detail. In other words, this is the minimum distance at
which two distinct points of a specimen can still be seen - either by
the observer or the microscope camera - as separate entities.

Resolution of bright field microscope is 0.2 micrometer.

Application of Bright field Microscope :-

Brightfield Microscope is used in several fields, from basic biology to

understanding cell structures in cell Biology, Microbiology, Bacteriology to
visualizing parasitic organisms in Parasitology.

Most of the specimens to viewed are stained using special staining to enable
visualization. Some of the staining techniques used include Negative staining
and Gram staining.

Some of its applications include:

 Used to visualize and study the animal cells.

 Used to visualize and study plant cells.
 Used to visualize and study the morphologies of bacterial cells.
 Used to identify parasitic protozoans such as Paramecium.

Advantages of bright field microscope :-

1. It is simple to use with few adjustments involved while viewing the

image.
2. It can be used to view both stained and unstained.
3. The optics of the microscope do not alter the color of the specimen.
4. The microscope can be adjusted and modified for better viewing such
as installing a camera, to form a digital microscope or in the way
image illumination is done such as by use of fluorochromes on the
specimen and viewing under a dark environment, forming a darkfield
microscope.
Disadvantages of bright field microscope :-

 The aperture diaphragm may cause great contrast which may distort
the outcome of the image, therefore iris diaphragm is preferred.
 It can not be used to view live specimens such as bacterial cells. Only
fixed specimens can be viewed under the brightfield microscope.
 Maximum magnification of the brightfield microscope is 100x but
modification can readjust the magnification to 1000x which is the
optimum magnification of bacterial cells.
 It has low contrast hence most specimens must be stained for them to
be visualized.
 Use of oil immersion may distort the image
 The use of coverslip may damage the specimen
 Staining may introduce extraneously unwanted details into the
specimen or contaminate the specimen.
 It is tedious to stain the specimen before visualizing it under the
brightfield microscope.
 The microscope needs a strong light source for magnification and
sometimes the light source may produce a lot of heat which may
damage or kill the specimen.