Rhodamine 123 Staining of Viable Cells

Another property of viable cells is the maintenance of electrochemical gradients across the plasma membrane. Functional subsets of this general phenomenon include the maintenance of pH and other ion gradients as well as the capacity for energy-yielding metabolism in mitochondria. These physiological processes can be exploited to distinguish viable from nonviable cells. One of the most commonly used probes for identifying viable cells is rhodamine 123, a cationic lipophilic dye that partitions into the low electrochemical potential of mitochondrial membranes. Active mitochondria in viable cells are stained bright green; loss of gradients within nonviable cells results in loss of the dye.

Additional Materials (also see Basic Protocol)

1 mg/ml rhodamine 123 (prepare fresh in distilled water) Cell suspension in culture medium appropriate for the cell type

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