A method for preparing quick‐frozen, freeze‐substituted cells for transmission electron microscopy and immunocytochemistry

R. G. Nagele; M. C. Kosciuk; S. M. Wang; D. A. Spero; H. Lee

doi:10.1111/j.1365-2818.1985.tb02645.x

A method for preparing quick‐frozen, freeze‐substituted cells for transmission electron microscopy and immunocytochemistry

R. G. Nagele, M. C. Kosciuk, S. M. Wang, D. A. Spero, H. Lee

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13 Scopus citations

Abstract

A quick‐freeze, freeze‐substitution method is described which employs glutaraldehyde as well as osmium tetroxide (OsO₄) in a ‘double‐fixation’ protocol comparable to that used for conventional transmission electron microscopy. Cultured cells are quick‐frozen in Freon 22 and freeze‐substituted in an ethanolic solution of glutaraldehyde. Specimens destined for TEM are postfixed in OsO₄ in acetone, embedded in Epon‐Araldite, and sectioned. This method yielded ultrastructural preservation which was comparable to that obtained from methods employing OsO₄ alone as a freeze‐substitution fixative. However, if glutaraldehyde is used alone as a freeze‐substitution fixative, specimens can be processed for immunocytochemistry without additional treatment with permeabilizing agents. 1985 Blackwell Science Ltd

Original language	English (US)
Pages (from-to)	291-301
Number of pages	11
Journal	Journal of Microscopy
Volume	139
Issue number	3
DOIs	https://doi.org/10.1111/j.1365-2818.1985.tb02645.x
State	Published - Sep 1985
Externally published	Yes

All Science Journal Classification (ASJC) codes

Pathology and Forensic Medicine
Histology

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10.1111/j.1365-2818.1985.tb02645.x

Cite this

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title = "A method for preparing quick‐frozen, freeze‐substituted cells for transmission electron microscopy and immunocytochemistry",

abstract = "A quick‐freeze, freeze‐substitution method is described which employs glutaraldehyde as well as osmium tetroxide (OsO4) in a {\textquoteleft}double‐fixation{\textquoteright} protocol comparable to that used for conventional transmission electron microscopy. Cultured cells are quick‐frozen in Freon 22 and freeze‐substituted in an ethanolic solution of glutaraldehyde. Specimens destined for TEM are postfixed in OsO4 in acetone, embedded in Epon‐Araldite, and sectioned. This method yielded ultrastructural preservation which was comparable to that obtained from methods employing OsO4 alone as a freeze‐substitution fixative. However, if glutaraldehyde is used alone as a freeze‐substitution fixative, specimens can be processed for immunocytochemistry without additional treatment with permeabilizing agents. 1985 Blackwell Science Ltd",

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AU - Nagele, R. G.

AU - Kosciuk, M. C.

AU - Wang, S. M.

AU - Spero, D. A.

AU - Lee, H.

PY - 1985/9

Y1 - 1985/9

N2 - A quick‐freeze, freeze‐substitution method is described which employs glutaraldehyde as well as osmium tetroxide (OsO4) in a ‘double‐fixation’ protocol comparable to that used for conventional transmission electron microscopy. Cultured cells are quick‐frozen in Freon 22 and freeze‐substituted in an ethanolic solution of glutaraldehyde. Specimens destined for TEM are postfixed in OsO4 in acetone, embedded in Epon‐Araldite, and sectioned. This method yielded ultrastructural preservation which was comparable to that obtained from methods employing OsO4 alone as a freeze‐substitution fixative. However, if glutaraldehyde is used alone as a freeze‐substitution fixative, specimens can be processed for immunocytochemistry without additional treatment with permeabilizing agents. 1985 Blackwell Science Ltd

AB - A quick‐freeze, freeze‐substitution method is described which employs glutaraldehyde as well as osmium tetroxide (OsO4) in a ‘double‐fixation’ protocol comparable to that used for conventional transmission electron microscopy. Cultured cells are quick‐frozen in Freon 22 and freeze‐substituted in an ethanolic solution of glutaraldehyde. Specimens destined for TEM are postfixed in OsO4 in acetone, embedded in Epon‐Araldite, and sectioned. This method yielded ultrastructural preservation which was comparable to that obtained from methods employing OsO4 alone as a freeze‐substitution fixative. However, if glutaraldehyde is used alone as a freeze‐substitution fixative, specimens can be processed for immunocytochemistry without additional treatment with permeabilizing agents. 1985 Blackwell Science Ltd

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A method for preparing quick‐frozen, freeze‐substituted cells for transmission electron microscopy and immunocytochemistry

Abstract

All Science Journal Classification (ASJC) codes

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