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Improved Methods For Preserving Macromolecular Structures And Visualizing Them By Fluorescence And Scanning Electron Microscopy, Paul B. Bell Jr., Barbara Safiejko-Mroczka
Improved Methods For Preserving Macromolecular Structures And Visualizing Them By Fluorescence And Scanning Electron Microscopy, Paul B. Bell Jr., Barbara Safiejko-Mroczka
Scanning Microscopy
To determine the optimal procedures to preserve cytoskeletal and other macromolecular structures for microscopic studies we have evaluated the effects of various methods to extract cultured cells. In this report, we compare results using different fixatives, crosslinking reagents, and permeabilization methods on (1) the labeling of cells for fluorescence microscopy with phalloidin or antibody against tubulin; and (2) the morphological preservation of macromolecular structures for scanning electron microscopy. Maximal labeling of F-actin with phalloidin was obtained by fixing cells in 4% para formaldehyde (PFA) and labeling the unextracted cells with methanolic phalloidin, whereas maximal labeling of tubulin required prefixation with …
The Effects Of Diethyldithiocarbamate (Ddc) On The Astrocytic Cytoskeleton, Mary F. Mcmanus, Louis D. Trombetta
The Effects Of Diethyldithiocarbamate (Ddc) On The Astrocytic Cytoskeleton, Mary F. Mcmanus, Louis D. Trombetta
Scanning Microscopy
The dithiocarbamates are a group of compounds that are used extensively in industry, agriculture and medicine. Exposure to these compounds has caused deleterious effects to both the central and peripheral nervous systems. Cultured rat hippocampal astroglia treated with 35 μg/ml diethyldithiocarbamate (DDC) in media were studied for alterations to the cytoskeleton. Examination by both immunohistochemistry and scanning electron microscopy revealed disruption of the cytoskeletal elements. This occurred in a progressive time-dependent manner. Electrophoretic patterns demonstrated two cytoskeletal protein alterations. The microtubular protein, β-tubulin, appeared to have an altered mobility while the major intermediate filament protein, glial fibrillary acidic protein (GF …