Electron microscopic studies of the endoplasmic reticulum in whole-mount cultured cells fixed with potassium...

A method for visualizing the endoplasmic reticulum and other membrane organelles in whole-mount cells with a standard, 60-kV transmission electron microscope has been developed. By use of a new formulation of potassium permanganate as a fixative, intracellular membranes were preserved and stained, while cytosolic proteins were digested, giving a pattern of membranous organelles against a clear background, suitable for transmission EM of whole-mount cells at 60 kV. Mitochondria, lysosomes, and ER were clearly visible in whole-mount cells fixed by this method. We have employed this technique to examine the organization of the ER in a variety of different cell lines. This method also allowed visualization of the three-dimensional organization, relationships, and fine structure of mitochondria. With prolonged permanganate fixation, mitochondrial cristae were clearly visible in whole-mount cells. This method was also useful for fixation and staining of thin sections, and allowed examination of thicker sections than previously possible, thus giving improved imaging of organelle relationships and fine structure. Using this method, we have examined the ER, mitochondria, and Golgi in thin section.