Immunophenotyping is a commonly used flow cytometry method for discriminating between different cell subsets based on intracellular and cell surface markers. A critical parameter for running a successful immunophenotyping assay is to ensure that you are analyzing live cells. Dead cells are not useful for data analysis and can do strange things when stained.
Take heed of these warnings to protect your cells from the zombie apocalypse!
- Live or Dead??? Several different stains exist for discriminating live cells from dead cells. Be sure to use a stain that works with your optimized panel and follow the specific instructions for processing and analyzing your cells to correctly identify living cells.
- Death by centrifuge. Cell processing and flow cytometry staining involves numerous centrifugation steps to isolate cell populations and wash cells after various staining steps. Typically, only low speed centrifugation is needed as higher speed spinning can have deadly consequences.
- Cold Catastrophe. Cell staining for flow cytometry can be carried out at different temperatures depending on the staining protocol. Some steps may allow cells to be refrigerated for a brief period of time, but deep cold exposure can have a fatal final result.