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Apoptosis - Organelle Analysis
During apoptosis there is often a collapse of the mitochondrial membrane potential. This can be detected in a number of ways by flow cytometry. Two dyes in particular are useful - CMXRos (also known as MitoTracker Red from Molecular Probes) and LDS-751 (from Exciton).
CMXRos has a chloromethyl group which allows accumulation in active mitochondria. Live cells that have active mitchondria are able to take up CMXRos but in cells that are undergoing apoptosis, the mitochondrial membrane potential decreases which means less dye accumulates in the mitchondria leading to a decrease in fluorescence. In the example below, dead cells can be identified by their TO-PRO-3 iodide positivity whereas apoptotic cells are seen as a population that shows reduced CMXRos fluorescence.
Laser Dye Styryl-751 (LDS-751) is often used in no-lyse peripheral blood preparations to identify nucleated cells - it is taken up by live cells and stains DNA, although not quantitatively. It is also likely to stain active mitochondria and so, is useful in the detection of apoptosis in the same way CMXRos is. Here we see that in apoptosis there is reduced LDS fluorescence in apoptotic cells.
The following protocols are available:- Detection of apoptotic cells by CMXRos in PDF format or HTML format
- Detection of apoptotic cells by LDS-751 in PDF format or HTML format.
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