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

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.