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.
Key wordsImmunogold labeling cryofixation freeze-substitution mammalian cultured cells lowicryl resin potassium permanganate
- 9.Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature 241, 20–22.Google Scholar
- 11.Griffiths, G., ed. (1993) Particle markers for immunoelectron microscopy. Fine Structure Immunocytochemistry. Springer-Verlag, Berlin, 279–306.Google Scholar