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A preparation method for observing intracellular structures by scanning electron microscopy
Author(s) -
Tanaka Keiichi,
Mitsushima Akira
Publication year - 1984
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1984.tb00487.x
Subject(s) - osmium tetroxide , fixative , osmium , scanning electron microscope , electron microscope , fixation (population genetics) , intracellular , microscopy , chemistry , biophysics , acid phosphatase , cytoplasm , materials science , pathology , biochemistry , biology , optics , medicine , composite material , enzyme , physics , ruthenium , gene , catalysis
SUMMARY In order to observe intracellular structures by scanning electron microscopy, excess cytoplasmic matrix must be removed from the fractured surface of cells. Previously we reported an Osmium‐DMSO‐Osmium method devised for this purpose. This method is very effective in revealing intracellular structures, but requires osmium tetroxide for initial fixation with some consequent disadvantages. In the present study, a revised Osmium‐DMSO‐Osmium method is reported, in which an aldehyde mixture is used as the initial fixative instead of osmium tetroxide. As fixation is carried out by perfusion in this revised method, better preservation of fine structures is achieved than by the original method, especially in the central nervous tissue which tends to suffer from post‐mortem degeneration. Moreover this method can be applied to cytochemical studies of intracellular structures with a scanning electron microscope (SEM). In this study, acid phosphatase of lysosomes is demonstrated in a coloured SEM micrograph.