Abstract
Techniques such as whole mount (high-voltage) electron microscopy of cultured cells (frequently extracted with non-ionic detergents), rapid-freezing (followed either by freeze-substitution or deep etching and rotary-replication), and reversible embedment of tissue have furnished novel insights into cytoskeletal architecture. Another approach in the examination of cytoskeletal morphology has been the modification of routine fixation and thin-section methods. Minimizing osmium exposure and including tannic acid in the primary fixative has been found to improve cytoskeletal preservation. Maupin and Pollard developed a fixation method in which a non-ionic detergent is added to the fixative to facilitate penetration of tannic acid into the cytoplasm. The images obtained by tannic acid fixation are frequently very electron-dense, however, and it is often difficult to clearly visualize the cytoplasmic filamentous system. This study describes results of an attempt to extract background material and reduce the density resulting from tannic acid fixation by reducing the concentration of aldehydes in the primary fixative.
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