1942•Journal of Scientific InstrumentsOpen access

Phase-Contrast Microscopy

Cecil Reginald Burch, Jonathan Stock

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Abstract

Slight optical non-homogeneities in certain transparent specimens are usually rendered visible by putting the specimen out of focus. One then sees not the non-homogeneities themselves, but the focal spots and lines they produce. Zernike's method of phase-contrast illumination, on the other hand, provides the maximum contrast when the specimen is accurately in focus, so that what is seen bears a closer resemblance to what is "really there". A simple method of adapting an ordinary microscope for phase-contrast is described, together with the preparation of a test-slide to check its performance. The method has proved helpful in studies of the interaction between leucocytes and mobile organisms.

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Slight optical non-homogeneities in certain transparent specimens are usually rendered visible by putting the specimen out of focus. One then sees not the non-homogeneities themselves, but the focal spots and lines they produce. Zernike's method of phase-contrast illumination, on the other hand, provides the maximum contrast when the specimen is accurately in focus, so that what is seen bears a closer resemblance to what is "really there". A simple method of adapting an ordinary microscope for phase-contrast is described, together with the preparation of a test-slide to check its performance. The method has proved helpful in studies of the interaction between leucocytes and mobile organisms.

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Available abstract

Slight optical non-homogeneities in certain transparent specimens are usually rendered visible by putting the specimen out of focus. One then sees not the non-homogeneities themselves, but the focal spots and lines they produce. Zernike's method of phase-contrast illumination, on the other hand, provides the maximum contrast when the specimen is accurately in focus, so that what is seen bears a closer resemblance to what is "really there". A simple method of adapting an ordinary microscope for phase-contrast is described, together with the preparation of a test-slide to check its performance. The method has proved helpful in studies of the interaction between leucocytes and mobile organisms.

Key concepts: Phase contrast microscopy, Contrast (vision), Zernike polynomials, Focus (optics), Optics, Microscopy, Optical microscope, Microscope

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