Freeze-fracturing of monolayers (capillary layers) of cell, membranes and viruses: some technical considerations.
A novel hinged device for freeze-fracturing of cell monolayer 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 piecie of mica (with polylysine) and this is covered with another piece of mica, thin brass plate of 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 application are: negative staining of intramembranous protein particles; chemical or physical analyses of single membrane leaflets; identification of protein complexes by immunoelectron microscopy, etc.
D000256:Adenoviridae / Q000648:ultrastructure; D000818:Animals; D001436:Bacteriorhodopsins; D002462:Cell Membrane / Q000648:ultrastructure; D004926:Escherichia coli / Q000648:ultrastructure; D005614:Freeze Fracturing / Q000295:instrumentation / Q000379:methods*; D006217:Halobacterium / Q000648:ultrastructure; D008565:Membrane Proteins; D008566:Membranes / Q000648:ultrastructure*; D009975:Orthomyxoviridae / Q000648:ultrastructure
D008565:Membrane Proteins; D001436:Bacteriorhodopsins