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

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

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

A quick‐freeze, freeze‐substitution method is described which employs glutaraldehyde as well as osmium tetroxide (OsO 4 ) 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 OsO 4 in acetone, embedded in Epon‐Araldite, and sectioned. This method yielded ultrastructural preservation which was comparable to that obtained from methods employing OsO 4 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 languageEnglish (US)
Pages (from-to)291-301
Number of pages11
JournalJournal of Microscopy
Volume139
Issue number3
DOIs
StatePublished - Jan 1 1985
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology

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