A New 16-Color Flow Cytometry Panel Protocol for the Functional Phenotyping of Circulating Human Cytotoxic T Cells and NK Cells

Building a panel for the intracellular staining of immune cells could pose several challenges. A protocol published recently in Cell Press STAR Protocols takes these into account and provides a detailed workflow for efficiently phenotyping circulating human cytotoxic T cells and NK cells. These effector cells are relatively rare but play a key role in immune defense and are frequently investigated with respect to various diseases.

The protocol offers step-by-step procedures for sample preparation and cell staining and provides a carefully designed 16-color flow cytometry panel and gating strategy for identifying cytotoxic T cells. Various troubleshooting tips are also included.

The protocol helps in the intracellular analysis of granzyme K, granzyme B and perforin. This simplified workflow can also be used for the functional phenotyping of other circulating immune cell types. In addition, it has also simplified the process of lysis and fixation of nucleated red blood cells, which can then be used for surface marker analysis.

Check out the “Functional phenotyping of circulating cytotoxic T cells and NK cells using a 16-color flow cytometry panel” STAR protocol.