Research Group of
Johannes A. Schmid
JC-1 stain of apoptotic cells
JC-1 (5, 5´, 6, 6´-tetrachloro-1, 1´, 3, 3´-tetraethylbenzimidazol-carbocyanine iodide) is a lipophilic fluorescent cation that incorporates into the mitochondrial membrane, where it can form aggregates due to the physiological membrane potential of mitochondria. This aggregation changes the fluorescence properties of JC-1 leading to a shift from green to orange fluorescence. Intact living cells stained with JC-1 therefore exhibit a pronounced orange fluorescence of mitochondria, which is detectable by flow analysis (in the FL-2 channel). Apoptosis results in a break-down of the mitochondrial membrane potential and a subsequent decrease of the orange fluorescence (and a slight increase of the green fluorescence). By that means, apoptotic cells can be easily distinguished from non-apoptotic cells.
JC-1 is prepared as a 1000x stock solution in DMSO (5 mg/ml).
Approximate detection settings on FACSort:
FL1: 360 V (log)
other settings: cell type specific, e.g. for TF-1:
Example of an analysis: