Novel violet fluorescent nanoparticles for immunophenotyping using flow cytometry

Abstract

Abstract A novel series of fluorescent nanoparticles with violet excitation have been developed for use with flow cytometry antibodies. These exceptionally bright, photostable dyes also have narrow emission spectra, making them ideal for inclusion in high parameter multicolor panels. For this study, StarBright Violets 440, 515, 610, 670, 710 and 790 were conjugated to human B-cell, T-cell, and Treg marker monoclonal antibodies and used to immunophenotype human peripheral blood mononuclear cells (PBMCs) in an 8-color panel. Tests with this panel were conducted in different staining buffers and compared to a panel with dyes having similar spectra and the same clonality. The new StarBright multicolor panel showed better resolution of Treg populations and reliable identification of the different T- and B-cell populations across different stain buffers and conditions, which makes the StarBright Violet nanoparticles an excellent addition to the field of high parameter flow cytometry.