Abstract
Successful electron microscope cytochemistry invariably requires a consideration of the initial methods for tissue processing, particularly the fixation procedure. Preservation of enzyme activities usually requires a mild fixation, for instance, with formaldehyde or low concentrations of glutaraldehyde, because strong fixatives, such as osmium tetroxide, or high concentrations of glutaraldehyde usually inhibit enzyme activity. However, weak fixatives may lead to less optimal structural preservation, and a suitable balance has to be reached between adequate fixation and ultrastructural preservation. Therefore, the preparatory procedures may differ for different enzymes or for other substances. The choice of fixative and fixation procedure is crucial to the outcome of enzyme cytochemistry because enzymes vary considerably with respect to their sensitivity to different fixatives. Potassium-dependent p-nitrophenylphosphatase, which is a partial enzyme activity of Na,K-ATPase, can only be preserved with formaldehyde fixatives of low strength but is inhibited by glutaraldehyde fixation. In contrast, acid phosphatase is much more resistant and is active even after fixation in 3% glutaraldehyde for 2 hours.
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