Electron microscopy of free-floating cells: thin-layering technique, and selection of individual cells for ultramicrotomy.
A rapid and accurate procedure for electron microscopy of individual cells from suspensions (blood, peritoneal exudate, etc.) is described. After fixation of the sample with standard techniques, the particulate constituents are suspended in buffered 5% bovine serum albumin, thin-layered by gravity on clear supports (cover glasses or polyester slips) in which an orientation grid had been scored, and then immobilized by exposure of the preparations to acrolein vapors. The specimens are examined for cells of interest under a light microscope using interference or phase contrast; individual cells to be sectioned are documented in three photomicrographs taken at different magnifications. After this the specimens are embedded like ordinary cover slip preparations. When examining the face of the polymerized block under a light microscope, the position of the selected cell beneath the orientation grid relief can readily be relocated by the aid of the pre-embedding reference micrographs.
D000818:Animals; D001773:Blood Cells / Q000648:ultrastructure*; D056890:Eukaryota / Q000648:ultrastructure*; D006652:Histological Techniques; D006801:Humans; D008854:Microscopy, Electron; D008867:Microtomy