F
low cytometry is a binary system, where cells are either positive
or negative for a specific marker when using an appropriate
fluorescent dye-conjugated antibody to detect a specific antigen.
The parallel development of instrument hardware and dyes expands
the number of overall parameters that can be analyzed in a single
experimental run, with each additional parameter translating into
an exponential increase in data acquisition capacity and research
potential. Thanks to over 45 years of development, flow cytometry is
now an integral protein analysis tool that enables scientists to delve
progressively deeper into cellular characterization and profiling.
As the new millennium began, flow cytometry had reached a plateau,
as scientists could not detect more than 18 parameters simultaneously.
Around this time, Sirigen designed and optimized a novel class of dyes
derived from the chemistry behind Alan Heeger, Alan MacDiarmid,
and Hideki Shirakawa’s Nobel Prize-winning discovery of conductive
polymers. These new polymer-based dyes were groundbreaking and
led initially to a series of direct and energy transfer tandem dyes
excited by the violet laser.Researchers from BD Biosciences recognized the untapped potential
of the UV laser and decided to leverage Sirigen polymer dye
technology to create a new family of dyes excited by UV wavelength
lasers. Over three years, BD Biosciences developed seven BD Horizon
BrilliantTM Ultraviolet (BUV) Dyes. These dyes were designed and
optimized to work with the 355-nm UV laser rather than the 375-nmMaking the Most Out of the Ultraviolet Laser:
How BD Horizon Brilliant
TM
Ultraviolet Dyes Drive DiscoverySPONSORED CONTENT
Laser Fluorochrome
BV421BV480BV605BV786BB515PE PerCP-Cy5.5PE-Cy7APCViolet
405 nmBV421
BV480
BV605
BV786Blue
488 nmBB515
PE
PerCP-Cy5.5
PE-Cy7
Red
640 nm APCLaser Fluorochrome
BUV395BUV496BUV615BV421BV480BB515PE PE-Cy7APCUV
355 nmBUV395
BUV496
BUV615
Violet
405 nmBV421
BV480
Blue
488 nm BB515
Yellow-Green
561 nmPE
PE-Cy7
Red
640 nm APCLaser Marker FluorochromeViolet
405 nmCD24 BV421
CD27 BV480
CD19 BV605
CD20 BV786Blue
488 nmIgD BB515
CD38 PE
CD3 PerCP-Cy5.5
IgG PE-Cy7
Red
640 nm IgM APC9-Color Panel on a 5-Laser Instrument Spread Matrix
9-Color Panel on a 3-Laser Instrument Spread Matrix
Laser Marker FluorochromeUV
355 nmCD19 BUV395
CD3 BUV496
IgD BUV615Violet
405 nmCD24 BV421
CD20 BV480
Blue
488 nm CD27 BB515
Yellow-Green
561 nmCD38 PE
IgG PE-Cy7
Red
640 nm IgM APC
Figure 1: As compared to a 3-laser instrument configuration, designing a 9-color panel on a 5-laser instrument
is simplified by a more balanced fluorochrome distribution and reduced number of challenging fluorochrome
combinations, represented as yellow and red boxes in the spread matrices.
BD Horizon Brilliant™ Blue Reagents (BB)
BD Horizon Brilliant™ Ultraviolet Reagents (BUV)
BD Horizon Brilliant Violet™ Reagents (BV)