DOI: 10.1002/jemt.1060070302. Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and high‐resolution scanning and scanning transmission electron microscopy

Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and high‐resolution scanning and scanning transmission electron microscopy

10.1002/jemt.1060070302

Crossref journal-article

Wiley

Journal of Electron Microscopy Technique (311)

Abstract

AbstractThis paper describes the use of sputter coating to prepare detergent‐extracted cytoskeletons for observation by scanning (SEM), scanning transmission (STEM), inverted contrast STEM, and transmission (TEM) electron microscopy. Sputtered coats of 1–2 nm of platinum or tungsten provide both an adequate secondary electron signal for SEM and good contrast for STEM and TEM. At the same time, the grain size of the coating is sufficiently fine to be just at (platinum) or below (tungsten) the limit of resolution for SEM and STEM. In TEM, the granular structure of platinum coats is resolved, and platinum decoration artifacts are observed on the surface of structures. The platinum is deposited as small islands with a periodic distribution that may reveal information about the underlying molecular structure. This method produces samples that are similar in appearance to replicas prepared by low‐angle rotary shadowing with platinum and carbon. However, the sputter‐coating method is easier to use; more widely available to investigators; and compatible with SEM, STEM, and TEM. It may also be combined with immunogold and other labeling methods. While TEM provides the highest resolution images of sputter‐coated cytoskeletons, it also damages the specimens owing to heating in the beam. In SEM and STEM cytoskeletons are stable and the resolution is adequate to resolve individual microfilaments. The best single method for visualizing cytoskeletons is inverted contrast STEM, which images both the metal‐coated cytoskeletal structures and electron‐dense material within the nucleus and cytoplasm as white against a dark background. STEM and TEM were both suitable for visualizing colloidal gold particles in immunolabeled samples.

Bibliography

Bell, P. B., Lindroth, M., & Fredriksson, B. (1987). Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and highâresolution scanning and scanning transmission electron microscopy. Journal of Electron Microscopy Technique, 7(3), 149â159. Portico.

Authors 3

Paul B. Bell (first)
Margaretha Lindroth (additional)
Bengt‐Arne Fredriksson (additional)

References 18 Referenced 15

{'key': 'e_1_2_1_2_1', 'first-page': '139', 'article-title': 'The application of scanning electron microscopy to the study of the cytoskeleton of cells in culture', 'author': 'Bell P. B.', 'year': '1981', 'journal-title': 'Scanning Electron Microsc. 1981'} / Scanning Electron Microsc. 1981 / The application of scanning electron microscopy to the study of the cytoskeleton of cells in culture by Bell P. B. (1981)
{'key': 'e_1_2_1_3_1', 'first-page': '45', 'volume-title': 'Science of Biological Specimen Preparation 1983', 'author': 'Bell P. B.', 'year': '1984'} / Science of Biological Specimen Preparation 1983 by Bell P. B. (1984)
{'key': 'e_1_2_1_4_1', 'first-page': '351', 'article-title': 'Some factors affecting the preservation of the cytoskeleton of cells in culture for scanning electron microscopy', 'volume': '1983', 'author': 'Bell P. B.', 'year': '1983', 'journal-title': 'Scanning Electron Microsc.'} / Scanning Electron Microsc. / Some factors affecting the preservation of the cytoskeleton of cells in culture for scanning electron microscopy by Bell P. B. (1983)
Bell P. B. Rundquist I. Svensson I. andCollins V. P.(1987a) The use of cytofluorometry to evaluate the binding of antibodies to the cytoskeleton of cultured cells.J. Histochem. Cytochem. in press. (10.1177/35.12.3680931)
Bell P. B. Rundquist I. Svensson I. andCollins V. P.(1987b) Formaldehyde sensitivity of a GFAP epitope‐removed by extraction with high salt.J. Histochem. Cytochem. in press. (10.1177/35.12.2445810)
{'key': 'e_1_2_1_7_1', 'first-page': '135', 'article-title': 'Cultured human glial and glioma cells', 'volume': '24', 'author': 'Collins V. P.', 'year': '1983', 'journal-title': 'Int. Rev. Exp. Pathol.'} / Int. Rev. Exp. Pathol. / Cultured human glial and glioma cells by Collins V. P. (1983)
{'key': 'e_1_2_1_8_1', 'first-page': '401', 'article-title': 'Monoclonal antibodies to glial fibrillary acidic protein in the cytological diagnosis of brain tumors', 'volume': '28', 'author': 'Collins V. P.', 'year': '1984', 'journal-title': 'Acta Cytol. (Baltimore)'} / Acta Cytol. (Baltimore) / Monoclonal antibodies to glial fibrillary acidic protein in the cytological diagnosis of brain tumors by Collins V. P. (1984)
{'key': 'e_1_2_1_9_1', 'first-page': '269', 'article-title': 'Monoclonal antibodies to glial fibrillary acidic protein', 'volume': '91', 'author': 'Collins V. P.', 'year': '1983', 'journal-title': 'Acta. Pathol. Microbiol. Immunol. Scand. [A]'} / Acta. Pathol. Microbiol. Immunol. Scand. [A] / Monoclonal antibodies to glial fibrillary acidic protein by Collins V. P. (1983)
10.1083/jcb.98.6.1973
10.1016/0014-4827(79)90206-4
10.1083/jcb.86.1.212
10.1002/sca.4950090202
10.1007/BF00712117
{'key': 'e_1_2_1_15_1', 'first-page': '907', 'article-title': 'Design and operation of a high efficiency magnetron sputter coater', 'volume': '1982', 'author': 'Nockolds C. E.', 'year': '1982', 'journal-title': 'Scanning Electron Microsc.'} / Scanning Electron Microsc. / Design and operation of a high efficiency magnetron sputter coater by Nockolds C. E. (1982)
{'key': 'e_1_2_1_16_1', 'first-page': '55', 'volume-title': 'Cancer', 'author': 'Pontén J.', 'year': '1975'} / Cancer by Pontén J. (1975)
10.1111/j.1699-0463.1968.tb03502.x
10.1073/pnas.78.7.4329
{'key': 'e_1_2_1_19_1', 'first-page': '599', 'article-title': 'Organization of actin meshworks in cultured cells: The leading edge', 'volume': '66', 'author': 'Small J. V.', 'year': '1981', 'journal-title': 'Cold Spring Harbor Symp. Quant. Biol.'} / Cold Spring Harbor Symp. Quant. Biol. / Organization of actin meshworks in cultured cells: The leading edge by Small J. V. (1981)

Dates

Type	When
Created	20 years, 6 months ago (Feb. 24, 2005, 2:23 a.m.)
Deposited	1 year, 10 months ago (Oct. 21, 2023, 4:38 a.m.)
Indexed	11 months, 3 weeks ago (Sept. 6, 2024, 5:39 a.m.)
Issued	37 years, 9 months ago (Nov. 1, 1987)
Published	37 years, 9 months ago (Nov. 1, 1987)
Published Online	20 years, 6 months ago (Feb. 4, 2005)
Published Print	37 years, 9 months ago (Nov. 1, 1987)

Funders 0

None

BibTeX

@article{Bell_1987, title={Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and high‐resolution scanning and scanning transmission electron microscopy}, volume={7}, ISSN={1553-0817}, url={http://dx.doi.org/10.1002/jemt.1060070302}, DOI={10.1002/jemt.1060070302}, number={3}, journal={Journal of Electron Microscopy Technique}, publisher={Wiley}, author={Bell, Paul B. and Lindroth, Margaretha and Fredriksson, Bengt‐Arne}, year={1987}, month=nov, pages={149–159} }

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