PMID: 46631Jan 1, 1975

Use of peroxidatic-enzyme staining to enhance resolution of cultured mammalian cells under phase microscopy

Stain Technology
I I Singer

Abstract

Staining of glutaraldehyde-fixed mammalian cells with peroxidatic enzymes (horseradish peroxidase or horse heart cytochrome c) greatly enhances resolution of their structure under phase microscopy. The topography of cell processes and regions of intercellular contact and overlapping is resolved precisely, even in dense cultures mounted in media which ordinarily do not permit clear demonstration of these areas. The technique is therefore a useful aid to the study of cultured cells with phase optics. Labeling depends on introducing free aldehydes into cells through the use of bifunctional fixatives such as glutaraldehyde. Acetone or formaldehyde fixation prevents staining, and labeling intensity is greatly diminished by pretreatment with spermine, a polyamine that reacts with glutaraldehyde. Electron microscopy reveals that peroxidase tags membranes preferentially; some areas are labeled smoothly, others in a punctate manner. Ribosomes are sharply contrasted, but nuclei remain unstained. Cytochrome c labels condensed nuclear chromatin intensely, and also stains ribosomes and portions of the cytoplasmic ground substance; membranes are mostly unmarked.

Citations

Jul 1, 1972·The Histochemical Journal·H R Miller
Nov 1, 1969·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M J Karnovsky, D F Rice
Apr 1, 1966·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·R C Graham, M J Karnovsky
May 1, 1954·Experimental Cell Research·M ABERCROMBIE, J E HEAYSMAN

Related Concepts

Ovoperoxidase
CYCS
Tissue Membrane
Enzymes, antithrombotic
Peroxidases
Ground Substance
Cell Process Structure
Sample Fixation
Acetone Measurement
Nuclear Chromatin

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