Practical advice to set up and troubleshoot your flow cytometry experiment for immune monitoring.
Samples have High Background
High background usually indicates that the staining protocol needs optimization. Make sure the sample is of high-quality. Use a viability stain to stain dead and dying cells, as dead cells cause significant unspecific background staining, and focus your panel on live cells.
Then optimize by varying the incubation time. Keeping non-specific binding to a minimum during the optimization will help to boost the signal-to-noise ratio. Finally, thoroughly wash the stained sample to eliminate unbound reagents and use a blocking reagent, like Fc block, to manage persistent interference from non-specific molecular interactions.
Further Help with Troubleshooting Flow Cytometry
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MHC Dextramer® Reagents
Sensitively quantify target immune cells by flow cytometry with a strong signal-to-noise ratio.