At the beginning of the new millennium, researchers had reached a plateau in the number of parameters that could be analysed in a flow cytometry experiment. This was due to the limited number of fluorochromes available and the overlaps that occurred in their excitation spectra with the available lasers. Then the Nobel Prize-winning discovery of conducting polymers by Alan Heeger, Alan MacDiarmid and Hideki Shirakawa led to the design of a new generation of direct and energy transfer tandem dyes, which could be excited by the violet laser.
Realising the potential of these polymer dyes, researchers from BD Biosciences created a new family of dyes that could be excited by the UV laser. To avoid cross-excitation by the violet laser, the UV dyes were optimised to work with the 355 nm laser. Called the BD Horizon Brilliant™ Ultraviolet Dyes (BUV), the seven members of this family have dramatically expanded the number of markers that can be simultaneously investigated in a single tube assay.
By using the BUV dyes, researchers are now able to extend the usable emission spectrum in UV laser equipped flow cytometers. These dyes also permit greater flexibility in panel design and a reduced spillover by allowing for the distribution of fluorochromes across more lasers and a wider spectrum.
The parallel developments in instrument hardware, coupled with the development in dyes, have resulted in an exponential increase in data generation, acquisition capacity, and research potential. Researchers can now avail of an even more extensive portfolio of fluorochromes than before to build panels for their conventional flow cytometry experiments, yielding high content information and improved biological insights as a consequence. The BUV reagents also allow researchers to obtain better quality data in low parameter panels by enabling a more balanced distribution of fluorochromes across more lasers and allow for good resolution of poorly expressed markers.
This article in TheScientist® traces the evolution and the development of BUV dyes from the early days and explains how these dyes have now made the UV laser mainstream.