1987
DOI: 10.1002/sca.4950090202
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TEM‐, SEM‐ and STEM‐studies of sputter‐coated cytoskeletons

Abstract: A technique for sputter coating of cytoskeletons from detergent extracted cells is described. The method allows their study in high resolution TEM, SEM and STEM. The use of cytoskeletons is also a valuable model system for the evaluation of metal coating techniques, allowing studies of deposited metal on fine filamentous structures of varying thicknesses.Pt and W were sputtered at 1 to 5 nm, and comparisons were made with reference to contrast and granularity. Both metals gave good topographical contrast, but … Show more

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Cited by 8 publications
(17 citation statements)
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“…Preservation of all the major cytoskeletal components is in line with previous studies in which 0.1-0.5% Triton X-100, together with buffered or stabilizing buffers (Pudney & Singer, 1979;Henderson & Weber, 1979;Heuser & Kirschner, 1980;Schliwa & Blerkom, 1981;Bell et al, 1987Bell et al, , 1989Lindroth et al, 1987), or replica technique combined with detergentextraction (Heuser & Kirschner, 1980), were used.…”
Section: Composition Of the Detergent-extracted Whole-mount Cytoskelesupporting
confidence: 51%
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“…Preservation of all the major cytoskeletal components is in line with previous studies in which 0.1-0.5% Triton X-100, together with buffered or stabilizing buffers (Pudney & Singer, 1979;Henderson & Weber, 1979;Heuser & Kirschner, 1980;Schliwa & Blerkom, 1981;Bell et al, 1987Bell et al, , 1989Lindroth et al, 1987), or replica technique combined with detergentextraction (Heuser & Kirschner, 1980), were used.…”
Section: Composition Of the Detergent-extracted Whole-mount Cytoskelesupporting
confidence: 51%
“…Whole-mount electron microscopy is a widely used technique to study three-dimensional relationships of various cell components, such as nuclei, the plasma membrane and cytoskeletal structures (Heuser & Kirschner, 1980;Schliwa & Blerkom, 1981;Lindroth et al, 1987;Bell et al, 1987Bell et al, , 1989. In order to make wholemount preparations, cultured cells are usually treated with a detergent in an appropriate buffer to extract the soluble cytoplasmic components (Osbom & Weber, 1977).…”
Section: Introductionmentioning
confidence: 99%
“…Thus the platinum grain structure can be resolved in the microscope in both SEM and STEM modes, but it is not sharply focused. Tungsten coats of similar thickness do not show this grainy substructure in either scanning mode (Lindroth et al, 1987).…”
Section: Resultsmentioning
confidence: 97%
“…PBS-A: phosphate buffered saline without calcium or magnesium (137 mM NaC1, 3 mM KC1, 0.5 mM Na2HP04, 1.5 mM KH2P04, pH 7.4); PFA: paraformaldehyde, 20% aqueous stock solution diluted to 4% just prior to use. SB: microtubule-stabilizing buffer (1mM EGTA, 4% polyethylene glycol 6000,100 mM PIPES, pH 6.9) (Bell et al, 1987;Henderson and Weber, 1979). Thsb: 0.5% Triton in HSB.…”
Section: Reagentsmentioning
confidence: 99%
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