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Freeze‐fracturing of monolayers (capillary layers) of cells, membranes and viruses: some technical considerations
Author(s) -
Nermut M. V.,
Williams Lynn D.
Publication year - 1977
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1977.tb00023.x
Subject(s) - monolayer , membrane , mica , capillary action , materials science , adsorption , chemistry , nanotechnology , composite material , biochemistry , organic chemistry
SUMMARY A novel hinged device for freeze‐fracturing of cell monolayers in the Balzers freeze‐etch unit is described. It is economical on biological material and enables oriented adsorption of sheet‐like membrane fragments. For freeze‐fracturing ‘by hand’ a monolayer is formed on a positively charged piece of mica (with polylysine) and this is covered with another piece of mica, thin brass plate or filter paper. Such a sandwich is frozen in liquid nitrogen and fractured by means of forceps. Several modifications of this technique as well as practical examples are described. Among possible applications are: negative staining of intramembranous protein particles; chemical or physical analyses of single membrane leaflets; identification of protein complexes by immunoelectron microscopy, etc.