Expanding panel size in spectral flow cytometry with StarBright Dyes excitable by the ultraviolet (355 nm), violet (405 nm), blue (488 nm) and yellow (561 nm) lasers

Abstract The recently launched fluorescent dyes from Bio-Rad, StarBright Dyes, deliver tunable brightness and spectral properties, greater stability, improved lot-to-lot reproducibility, and spectral consistency. Specifically designed for multicolor flow cytometry with researchers needs in mind, StarBright Dyes address the common pain points in flow such as brightness, broad emission spectra, staining consistency, and ease-of-use, solving issues of signal resolution when constructing complex panels. These bright dyes have the additional benefit of a unique full spectral profile, making them ideal for inclusion in spectral flow cytometry, when creating multicolor panels. Here we present data of our newest additions to the StarBright range, the StarBright Blue and StarBright Yellow Dyes on a five-laser spectral cell analyzer. Both dye series are bright and allow expansion across the 488 and 561 lasers giving more dye choice for all panel sizes but are particularly useful when large panel design is required. In this study we show, when StarBright Blue and StarBright Yellow Dyes are combined with other members of the StarBright Dye range, polymer dyes and traditional fluorescent dyes, very large immunophenotyping panels can be constructed allowing identification of many peripheral blood subsets. In addition, novel dye combinations can be identified increasing the numbers of dyes and therefore markers identified from one sample. When combined with their ability to be fixed in both PFA based and alcohol-based fixatives, with minimal spectral changes, and to be pre-mixed for up to 33 days, the flexibility of StarBright Dyes means they are ideal for new and expanding existing panels regardless of the type of flow cytometer being used.