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  Phase Contrast Microscope

Phase Contrast Microscope:

Phase contrast Microscope first explained in the year 1934 by Dutch Physicist Frits Zernike, is a contrast-improving optical method that can be employed to generate high-contrast images of transparent specimens.

The total detail of living cells is undetectable in bright field microscope as there is too little contrast among structures with identical transparency. Such organisms are noticed with a phase contrast microscope. The Phase Contrast Microscope is broadly employed for observing biological specimens. Phase Contrast Microscope is a kind of light microscope that improves the contrasts of colorless and transparent objects by influencing the optical path of light. In phase contrast microscope, the light passing via a transparent portion of the specimen travels slower.

The condenser of a phase contrast microscope contains an annular stop, an opaque disk with a thin transparent ring, that generates a hollow cone of light (figure is as shown).

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                                       Figure: The light pathway of Phase Contrast Microscopy

Whenever this cone of light passes via a cell, similar light rays are bent due to density variation and refractive index in the specimen. The deviated light is focused to build up an image of the object. Undeviated light rays hit a phase ring in the phase plate, located in the objective. Therefore in phase contrast microscope one set of light rays comes directly from the light source, and the other set of light comes from the reflected or diffracted from the specific structure of the specimen. Whenever two set of light rays are brought altogether they form an image of the specimen on the ocular lens, having regions that are relating light (i.e., phase) via the shades of gray to black. The interior structure of the specimen becomes more sharply stated in phase contrast microscopy.

 

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