How do I discriminate between live cells and dead cells when doing a flow cytometry analysis?
Discrimination between viable and non-viable cells can be carried out with the use of the 7-AAD viability dye (eBioscience product 00-6993). Add 5 l per million cells of this dye in the samples and incubate for 5 minutes before analysis on the flow cytometer. The 7-AAD will mark the non-viable cells by binding to the nuclei of those cells. The nucleic acid of viable cells will not be accessible to the dye and will not be stained. When analyzing the data collected, gate out all cells stained with the viability dye observed typically in the third channel of the 488nm laser.
Discrimination between viable and non-viable cells can be carried out with the use of the 7-AAD viability dye (cat. 00-6993) or Propidium Iodide (PI) Staining Solution (cat. 00-6990). The 7-AAD or PI will mark the non-viable cells by binding to the nuclei of those cells. The nucleic acid of viable cells will not be accessible to the dye and will not be stained. When analyzing the data collected, gate out all cells stained with the viability dye. When staining for intracellular proteins, use the Fixable Viability Dye eFluor 450, 660, or 780. For more information, please see our Functional Dyes and Reagents webpage.
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