Immunoelectron Microscopy of Cryofixed Freeze-Substituted Mammalian Tissue Culture Cells

Mammalian tissue cultured cells are widely used in cell biology research. Immunoelectron microscopy is a powerful technique to define the subcellular localization of targeted antigens in the cultured cells. Cryofixation is now generally accepted as the best initial fixation step, to preserve the cellular fine structures and antigenicity. This chapter covers the practical procedures for immunoelectron microscopy of cryofixed freeze-substituted mammalian tissue cultured cells processed by high-pressure freezing.