Bright field microscopy

The name "brightIield" is derived Irom the Iact that the specimen is dark and contrasted by the
surrounding bright viewing Iield. Simple light microscopes are sometimes reIerred to as bright Iield
microscopes. It is the most elementary Iorm oI
microscope illumination techniques and is generally
used with compound microscopes.
!rinciple
In bright Iield microscopy a specimen is placed on
the stage oI the microscope and incandescent light
Irom the microscope`s 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 Iocus 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 magniIies the light and transmits it to an oracular lens or
eyepiece and into the user`s eyes. Some oI the light is absorbed by stains, pigmentation, or dense
areas oI 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 magniIications and lower light levels Ior
lower magniIications.
Light path of a bright field microscope
The light path oI a bright Iield microscope is extremely simple , no additional components are
required beyond the normal light microscope setup. The light path thereIore consists oI:
O Transillumination light source, commonly a halogen lamp in the microscope stand.
O ondenser lens which Iocuses light Irom the light source onto the sample.
O bjective lens which collects light Irom the sample and magniIies the image.
O culars and/or a camera to view the sample image
ommon components
The bright Iield microscope is made up oI several parts. They are:
O 1he base the structure that supports the unit and contains the control Ior the adjustment
oI light intensity, which is usually an illuminator or electrical source oI light.
O 72 the structure that provides support Ior the base and the lens system.

O bjective lenses these lenses are located nearest the object or specimen and enlarge the
image using a combination oI diIIerent magniIication powers. The objective lens
provides the aerial or primary image oI the specimen.
O cula7s these are the lenses that magniIy an image. These lenses are located nearest to
the viewer. The ocular lens magniIies the image to put the primary image in Iocus. This
helps provide the virtual or Iinal image, which is the product oI the ocular and objective
lenses.
O #evolving nosepiece allows rotation oI the objective lenses Ior better viewing position.
O $tage the platIorm where the microscope slide is laid and viewed.
O /just2ent knobs consist oI the coarse adjustment and the Iine adjustment knobs. The
coarse adjustment Iocuses the image using the 4X and 10X objective lenses. The Iine
adjustment lenses Iocus the image using the high dry and oil immersion objective lenses
(40X and 100X).
O on/ense7 this may be raised or lowered to Iocus the light Ior better viewing oI the
specimen.
dvantages
O rightIield microscopy is very simple to use with Iewer adjustments needed to be made
to view specimens.
O Some specimens can be viewed without staining and the optics used in the brightIield
technique don`t alter the color oI the specimen.
O It is adaptable with new technology and optional pieces oI equipment can be
implemented with brightIield illumination to give versatility in the tasks it can perIorm.
O For images that require viewing enhancements, a bright Iield microscope may be adjusted
manually, allowing manipulation oI the light source through the condenser, eIIectively
reducing the resolution.
O !olarising Iilters may also be used with the light source to help enhance specimen
Ieatures that are not apparent when under a white light.
O They also perIorm excellently Ior magniIication oI transparent, opaque or semi-
transparent specimens.
Limitations of Brightfield Microscopes
O rightIield microscopy can`t be used to observe living specimens oI bacteria, although
when using Iixed specimens, bacteria have an optimum viewing magniIication oI 1000x.
O It have low magniIication power and requires a strong light source Ior high magniIication
applications and intense lighting can produce heat that will damage specimens or kill
living microorganisms.
O The bright Iield microscope is advantageous Ior images that are either dark or those
which have signiIicant reIraction because the white background can eIIectively enhance
the image. For other images, a bright Iield microscope may not provide the best view.
O The resolution oI diIIraction is limited to about 0.2 micrometer. Lights that are out oI
Iocus located in the external area oI the Iocal plane Iurther reduce the clarity oI the
image. It have low resolving power.
O Some images also do not have the natural contrast that allows them to be studied and
viewed successIully using the bright Iield microscope. Some live specimens may also
prove diIIicult to view using a bright Iield microscope, limiting imaging in real time.
O some specimens have to be stained in order to provide or increase the contrast oI the
diIIerent structural details. e that as it may, staining may also introduce certain details
into the specimen that are not part oI the specimen itselI, but are instead a caused by the
processing. The apparent details may be misconstrued as a Ieature when in truth, they are
not. However, background illumination may be corrected when using the device.

&ses of the bright field microscope

rightIield Microscopes provide images that appear dark-colored against a bright Iield. They are
used extensively in classroom settings Ior the examination oI specimen in biology and chemistry.
This type oI microscope may be used Ior either stained or unstained specimens. In microbiology,
Ior example, simple stains such as crystal violet and methylene blue as well as diIIerential stains
such as Ilagellar stains, negative stains and endospore stains are oIten used to enhance details in
a specimen Ior observation. Images oI microorganisms may be magniIied to a maximum oI
400X. When specimens are stained, they can be Iurther magniIied with an oil immersion lens to
a maximum oI 1200X. rightIield Microscopes are also eIIective in bacteriological microscopy
because the oil immersion lens allows Ior increase in resolving power.