Principles of Phase Contrast Microscopy

Differences in light absorption are often negligible between living cells and their surrounding nutrient medium, as well as between the various intracellular components and plasma membranes, rendering these entities barely visible when observed by brightfield illumination. Phase contrast microscopy takes advantage of minute refractive index differences within cellular components and between unstained cells and their surrounding aqueous medium to produce contrast in these and similar transparent specimens.
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Contributing Authors

Douglas B. Murphy - Department of Cell Biology and Anatomy and Microscope Facility, Johns Hopkins University School of Medicine, 725 N. Wolfe Street, 107 WBSB, Baltimore, Maryland 21205.

Ron Oldfield - Department of Biological Sciences, Division of Environmental and Life Sciences, Macquarie University, New South Wales 2109, Australia.

Stanley Schwartz - Bioscience Department, Nikon Instruments, Inc., 1300 Walt Whitman Road, Melville, New York 11747.

Greenfield Sluder - Department of Cell Biology, University of Massachusetts Medical School, 377 Plantation Street, Worcester, Massachusetts 01605.

Tatsuro Otaki - Optical Design Department, Instruments Company, Nikon Corporation, 1-6-3 Nishi-Ohi, Shinagawa-ku, Tokyo, 140-8601, Japan.

Matthew Parry-Hill, Robert T. Sutter, Cynthia D. Kelly, Shannon H. Neaves, Omar Alvarado, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.