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Phtomicroghraph

The document outlines the basic principles of photomicrography, emphasizing the use of bright field and cross polarized illumination techniques in microscopy. It details the configuration of microscopes for photomicrography, including the placement of polarizing elements and the attachment of cameras. Finally, it describes the process of capturing photomicrographs by achieving maximum extinction through proper alignment of the polarizer and analyser.

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22 views4 pages

Phtomicroghraph

The document outlines the basic principles of photomicrography, emphasizing the use of bright field and cross polarized illumination techniques in microscopy. It details the configuration of microscopes for photomicrography, including the placement of polarizing elements and the attachment of cameras. Finally, it describes the process of capturing photomicrographs by achieving maximum extinction through proper alignment of the polarizer and analyser.

Uploaded by

Riya
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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Principle # 1.

Basic Principle:
• The basic principle of photomicrography involves the use of classical
microscopy techniques of bright field and cross polarized illumination. Most
microscopes used in biological laboratories are of transmitted light variety
and operate in the bright field mode. Placing a polarizing element into the
light path restricts the passage of light thus reducing the amount of
transmitted light to approximately 30% of the emitted value.
• To obtain cross polarized illumination from bright field microscope, two
individual polarizing elements (one is called polarizer and the other one
analyser) are inserted into the light path with their vector proportion planes
crossed at a 90° angle with respect to each other.
• When a sample is placed in the light path between the polarizer and the
analyser (i.e., crossed polarizers), the only light emitted will be that which is
refracted by the sample until it can pass unimpeded through the analyser.
Principle # 2. Microscope Configuration for Photo­micrography:
• The commonly used bright field micro­scope in which an external light
source is reflected into the sub-stage condenser through a mirror, can
easily be converted for use with polarizing elements (Fig. 15.6).
• The polarizer responsible for polarizing the light is taped onto the bottom
of the condenser. The analyser (the second polarizer) is inserted inside
the body of the microscope between the main body tube and the
eyepiece tube.
• Attaching a camera to the microscope is the last step. Microscope viewing
heads come in three varieties: monocular (one eyepiece), binocular (two
eyepieces) and trinocular (two eye­pieces and a photography lube).
• A camera can be adapted to each of these viewing heads. A simple
camera back will be sufficient for photomicrography as the camera is
needed only to store, expose, and advance the film. The microscope itself
acts as a camera lens.
Principle # 3. Taking the Photomicrograph:
• After both the polarizer and the analyser are in place and the
camera is fitted on the microscope, the specimen is placed on
the stage and is viewed directly into the eyepiece. The image of
the specimen is brought into focus and the polarizer is rotated
until the view-field becomes very dark (maximum extinction).
• At this point, the polarization direction is perpendicular between
the polarizer and analyser i.e. they are in the position of cross
polarizers as described above. This results in cross polarized
illumination which is needed for photomicrography. Now the
photomicrograph of the specimen is taken with the help of
camera already adapted to viewing head of the microscope.

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