Visualization of fission yeast cells by transmission electron microscopy

Abstract

This chapter deals with the preparation of fission yeast (Schizosaccharomyces) cells for ultrastructural examination. The structure of the cell must be preserved as close to the in vivo situation as possible. This can be achieved by either chemical or cryofixation; the latter will not be dealt with in this chapter. Aldehydes that cross-link proteins and permanganates that besides cross-linking also stain membranous and cell wall structures are used for chemical fixation. This step is followed by dehydration and embedding of the cells in epoxy or acrylic resin. Sectioning of the embedded material produces slices of the cells that have to be stained with heavy metals to increase contrast differences between different structures or can be used for immunodetection of antigens (polysaccharides or proteins) with specific primary antibodies and goldconjugated secondary antibodies.