Protocol for in-plate staining of clinical peripheral blood mononuclear cell samples for high-throughput analysis via spectral flow cytometry

Clinical trial research relies on robust assays to monitor immune responses in patients. We present a protocol for optimization of in-plate staining for spectral flow cytometry tailored for high-throughput analysis of peripheral blood mononuclear cell (PBMC) samples from clinical research correlative studies. We describe steps for antibody titration, optimal in-plate staining, data acquisition on Sony ID7000 spectral cell analyzer, and validation of a flow cytometry assay.