Abstract
Since the introduction of tannic acid as an additive in glutaraldehyde fixatives for electron microscopy, there have been numerous reports of ultrastructural findings that were not detectable after fixation without tannic acid. We have used tannic acid in studies on the spindle apparatus in insect spermatocytes to show that microtubules of the spindle are composed of 13 protofilaments. Tannic acid accumulates at the periphery of subunits of microtubules and with osmium stains those regions, leaving the subunits unstained. Hence, the subunits appear negatively stained.We have found that in addition to microtubules, other filamentous structures in cells appear negatively stained after fixation with glutaraldehyde-tannic acid. The most noteable are actin filaments in muscle cells. We report here results obtained from mouse striated muscle.Segments of gastrocnemius and vastus lateralis muscles are dissected in 3% glutaraldehyde in 0.1 M cacodylate buffer and subsequently fixed for 1 hr in the above fixative containing tannic acid (Mallinckrodt) in concentrations of 4 and 8%. Both concentrations gave similar results.
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Schedule a callMore From: Proceedings, annual meeting, Electron Microscopy Society of America
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