DOI: 10.1046/j.1365-2818.2003.01231.x. The use of filter membranes for high‐pressure freezing of cell monolayers

The use of filter membranes for high‐pressure freezing of cell monolayers

10.1046/j.1365-2818.2003.01231.x

Crossref journal-article

Wiley

Journal of Microscopy (311)

Abstract

SummaryRapid freezing of cells and tissues, followed by freeze‐substitution fixation and plastic embedding, has become a highly reliable method for preparing samples for imaging in the electron microscope. High‐pressure freezing is an efficient means of immobilizing suspensions of yeasts, thick pellets of mammalian cells, or small (< 0.5 mm) pieces of plant or animal tissue. Monolayers of cultured mammalian cells that are too thick for efficient immobilization by other modes of rapid freezing have also been successfully preserved by this method. Monolayer cultures are often important because they can be imaged by light microscopy (LM) both before and after their preparation for electron microscopy (EM). Additionally, some monolayer cultures serve as model systems for physiological processes, so it is important that cells under study can grow on a substrate that is both physiologically appropriate and convenient for EM processing. Here we describe a reliable method for preparing mammalian cell monolayers (PtK1 and polarized MDCK) for EM. Our protocol results in good preservation of cellular ultrastructure, it is a useful companion to studies of cell physioloy and, with some limitation, is suitable for correlative LM and EM.

Bibliography

Morphew, M. K., & Mcintosh, J. R. (2003). The use of filter membranes for highâpressure freezing of cell monolayers. Journal of Microscopy, 212(1), 21â25. Portico.

Authors 2

M. K. Morphew (first)
J. R. Mcintosh (additional)

References 19 Referenced 27

10.1083/jcb.99.5.1655
10.1002/jemt.1060130305
{'key': 'e_1_2_6_4_1', 'first-page': '47', 'article-title': 'Pep1p, a SCP110‐related calmodulin target at the centrosome of the fission yeast Schizosaccharomyces pombe', 'volume': '13', 'author': 'Flory M.', 'year': '2002', 'journal-title': 'Cell Growth Differentiation'} / Cell Growth Differentiation / Pep1p, a SCP110‐related calmodulin target at the centrosome of the fission yeast Schizosaccharomyces pombe by Flory M. (2002)
10.1002/jemt.1060030206
10.1006/jsbi.2000.4230
10.1111/j.1365-2818.1994.tb04785.x
10.1083/jcb.144.6.1135
10.1006/jsbi.1997.3919
10.1007/s004120050320
{'key': 'e_1_2_6_11_1', 'first-page': '175', 'volume-title': 'Science of Biological Specimen Preparation', 'author': 'Moor M.', 'year': '1987'} / Science of Biological Specimen Preparation by Moor M. (1987)
{'key': 'e_1_2_6_12_1', 'first-page': '131', 'volume-title': 'The Science of Biological Specimen Preparation', 'author': 'Müller M.', 'year': '1984'} / The Science of Biological Specimen Preparation by Müller M. (1984)
10.1006/jsbi.1998.3971
10.1111/j.1365-2818.1983.tb04200.x
10.1177/33.2.3881520
10.1046/j.1365-2818.2002.01085.x
{'key': 'e_1_2_6_17_1', 'first-page': '253', 'article-title': 'High Pressure freezing comes of age', 'volume': '3', 'author': 'Studer D.', 'year': '1989', 'journal-title': 'Scanning Microsc. Suppl'} / Scanning Microsc. Suppl / High Pressure freezing comes of age by Studer D. (1989)
10.1083/jcb.105.4.1623
{'key': 'e_1_2_6_19_1', 'first-page': '5', 'article-title': 'Cryofixation and cryosubstitution: a useful alternative in the analysis of cellular fine structure', 'volume': '37', 'author': 'Von Schack M.L.', 'year': '1993', 'journal-title': 'Eur. J. Histochem'} / Eur. J. Histochem / Cryofixation and cryosubstitution: a useful alternative in the analysis of cellular fine structure by Von Schack M.L. (1993)
10.1083/jcb.146.2.415

Dates

Type	When
Created	21 years, 10 months ago (Oct. 14, 2003, 6:34 a.m.)
Deposited	1 year, 10 months ago (Oct. 14, 2023, 1:19 p.m.)
Indexed	11 months, 2 weeks ago (Sept. 6, 2024, 8:38 p.m.)
Issued	21 years, 10 months ago (Oct. 1, 2003)
Published	21 years, 10 months ago (Oct. 1, 2003)
Published Online	21 years, 10 months ago (Oct. 1, 2003)
Published Print	21 years, 10 months ago (Oct. 1, 2003)

Funders 0

None

BibTeX

@article{Morphew_2003, title={The use of filter membranes for high‐pressure freezing of cell monolayers}, volume={212}, ISSN={1365-2818}, url={http://dx.doi.org/10.1046/j.1365-2818.2003.01231.x}, DOI={10.1046/j.1365-2818.2003.01231.x}, number={1}, journal={Journal of Microscopy}, publisher={Wiley}, author={Morphew, M. K. and Mcintosh, J. R.}, year={2003}, month=oct, pages={21–25} }

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