Rapid-freezing cytochemistry: preservation of tubular lysosomes and enzyme activity.
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.[1]References
- Rapid-freezing cytochemistry: preservation of tubular lysosomes and enzyme activity. Robinson, J.M., Karnovsky, M.J. J. Histochem. Cytochem. (1991) [Pubmed]
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