Brilliant Violet 421™ and Brilliant Violet 570™ antibody conjugates deliver consistent and superior performance on violet-laser equipped flow cytometers with varied specifications (65.4)

Abstract

Abstract Brilliant Violet™ derived fluorophores are novel and highly sensitive polymeric molecules that fluoresce upon excitation by a 405 nm laser line. In this study, Brilliant Violet 421™ and Brilliant Violet 570™ antibody conjugates were evaluated for their performance on violet laser equipped flow cytometers with varying instrument configurations. Instrument specific variables included sheath pressure, laser power, PMT gain and position, optical configuration, and filter sets. Cross beam spectral overlap was also evaluated for 488nm and 561nm lasers. All instrument configurations revealed significantly increased signal to noise ratio with Brilliant Violet™ as compared to traditional violet laser dyes. Brilliant Violet 421™ yielded logarithmic improvements in signal to noise ratio, while maintaining manageable spectral compensation properties. Brilliant Violet 421™ antibody conjugates against high and low-density surface markers also increased resolution and accuracy, notably for human CD25, CD56, CD127 and mouse NK1.1 and CD4. The Brilliant Violet™ conjugates were also found to be effective for detection of intracellular cytokines. Overall, Brilliant Violet™ 421 and Brilliant Violet™ 570 were determined to outperform traditional violet-excitable organic dyes on flow cytometers with a variety of specifications and settings.