Freeze fracture through the cytoskeleton, nucleus and nuclear matrix of lymphocytes studied by scanning electron microscopy

Abstract

The technique of delaying fixation until after freeze‐fracture and thawing, described in an earlier paper (Haggis & Bond, 1979), has been developed further for study of cells in culture, principally mouse lymphocytes stimulated by concanavalin A. Using a thin layer of cells, a cryoprotectant concentration of either 10% glycerol or dimethylsulphoxide, is sufficient to give good structural preservation after rapid freezing and thawing. Nuclear matrices and Triton‐permeabilized cells have been prepared from stimulated lymphocytes for comparative study. Polylysine‐coated fibrin support films have been found to provide a convenient means of handling cells and subcellular preparations during freeze fracture, critical point drying and mounting for high‐resolution scanning electron microscopy.