Conventional and Immunoelectron Microscopy of Mitochondria

Electron microscopy (EM) has been a central tool in delineating the subcellular organization and function of the eukaryotic cell. It has provided valuable information on the organization of the Golgi complex; the polarized distribution of proteins on the plasma membrane; and fundamental insights into the essential structure and function of mitochondria beginning with the first EM observations of Claude and Fullam on isolated mitochondria in 1944. Most significant for this volume is the contribution immunoelectron microscopy (IEM) has made in the study of mitochondrial dynamics and in demonstrating the localizations of key mitochondrial proteins in yeast, including, though not limited to, Dnm1p, Fiz1p, and Mgm1p. This chapter is not intended to provide a comprehensive review of all EM and IEM methods as there are a number of excellent books and reviews already available on these topics. Rather, this chapter provides detailed protocols of conventional EM and IEM methods successfully utilized in our center for the examination and analysis of mitochondria in yeast and mammalian cells.