High-Dimensional Immunophenotyping of Human Leukocytes by Spectral Cytometry

Spectral cytometry has rapidly evolved over the past two decades. The development of complete spectral analysis using the flow cytometer as a spectrometer has been actualized due to advances in optics, detectors, and electronics that have made single-cell flow spectroscopy a reality. Polychromatic flow cytometry is a complex process, including the design of antibody panels and instrument compensation. Spectral cytometry has contributed significantly to effectively dealing with the complexity of polychromatic flow cytometry. It has substantial potential to improve primary and clinical research. Here, we provide an overview of the methodology to develop a 15-color human polychromatic flow cytometry panel using spectral flow cytometry.