Bright field Microscopy

Introduction:

It is the most elementary form of microscope illumination techniques and is generally used
with compound microscopes.

The name "bright field" is derived from the fact that the specimen is dark and contrasted by
the surrounding bright viewing field. Simple light microscopes are sometimes referred to as
bright field microscopes.

How it works

In bright field microscopy a specimen is placed on the stage of the microscope and
incandescent light from the microscopes light source is aimed at a lens beneath the
specimen. This lens is called a condenser.

The condenser usually contains an aperture diaphragm to control and focus light on the
specimen; light passes through the specimen and then is collected by an objective lens
situated in a turret above the stage.

The objective magnifies the light and transmits it to an oracular lens or eyepiece and into the
users eyes. Some of the light is absorbed by stains, pigmentation, or dense areas of the
sample and this contrast allows you to see the specimen.

For good results with this microscopic technique, the microscope should have a light source
that can provide intense illumination necessary at high magnifications and lower light levels
for lower magnifications.

Uses and Advancements

To some extent, bright field microscopy is used in most disciplines requiring microscopic
investigation.

Because it is a simple method, this is the first type of microscopy students learn in schools.

The life sciences, particularly microbiology and bacteriology, have always relied on the
brightfield technique.
This technique can be used to view fixed specimens or live cells. Since many organic
specimens are transparent or opaque, staining is required to cause the contrast that allows
them to be visible under the microscope.

Different stains and staining techniques are used depending upon the type of specimen and
cell structure being examined. For example

Fuchsin is used to stain smooth muscle cells

Methylene blue is used to stain cell nuclei
Gram stain is used on bacteria and gives rise to the name gram-negative or gram-
positive bacteria based on the reaction of the bacteria to the stain. In fact, many
scientific journals will not accept microbiological research for publication that is not
supported by gram staining and brightfield illumination methodology. Most routine
medical microscopic examination of blood and tissue is performed using this
illumination technique.

Different complimentary techniques can be used to augment brightfield microscopy. By using

a polarizing filter this illumination technique can be used in geological microscopic research
and will reveal details not visible using white light.

Properly stained, microorganisms may be magnified to 1200x; utilizing an oil immersion

objective will increase resolution at this high magnification.

Advantages

Bright field microscopy is very simple to use with fewer adjustments needed to be made to
view specimens.

Some specimens can be viewed without staining and the optics used in the bright field
technique doesnt alter the color of the specimen.

It is adaptable with new technology and optional pieces of equipment can be implemented
with brightfield illumination to give versatility in the tasks it can perform.

Disadvantages

Certain disadvantages are inherent in any optical imaging technique.

 By using an aperture diaphragm for contrast, past a certain point, greater contrast adds
distortion. However, employing an iris diaphragm will help compensate for this
problem.
Bright field microscopy cant be used to observe living specimens of bacteria,
although when using fixed specimens, bacteria have an optimum viewing
magnification of 1000x.

Bright field microscopy has very low contrast and most cells absolutely have to be stained to
be seen; staining may introduce extraneous details into the specimen that should not be
present.

Also, the user will need to be knowledgeable in proper staining techniques.

Lastly, this method requires a strong light source for high magnification applications and
intense lighting can produce heat that will damage specimens or kill living microorganisms.