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Rapid-freezing cytochemistry: preservation of tubular lysosomes and enzyme activity

JM Robinson and MJ Karnovsky

Department of Cell Biology, Neurobiology, and Anatomy, Ohio State University, Columbus 43210.

We show that tubular structures present in phorbol ester-stimulated macrophages are sensitive to commonly used chemical fixatives (i.e., they usually become fragmented during fixation). These structures are well preserved in macrophages that are physically fixed by rapid- freezing and subsequent freeze-substitution in osmium-acetone. We have developed methods that combine rapid-freezing, freeze-substitution, and enzyme cytochemistry for preservation of these tubular structures and for detection of endocytosed material (i.e., horseradish peroxidase). This method of rapid-freeze cytochemistry may be useful in other situations where chemical fixation does not adequately preserve cell structures, particularly of membrane compartments.

Volume 39, Issue 6, pp. 787-792, 06/01/1991
Copyright © 1991 by The Histochemical Society


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