Lasers and Dyes

BD Horizon Brilliant™ Ultraviolet 563 (BUV563) (Ex Max 350 nm/Em Max 564 nm) is a tandem fluorochrome that combines BD Horizon™ BUV395 and an acceptor dye with an Em Max at 564 nm. Due to the excitation of the acceptor dye by other laser lines, there may be spillover into channels detecting PE (e.g., 575/26 nm filter) and PE-CF594 (e.g., 610/20 nm filter). BUV563 has been exclusively developed by BD Biosciences for instruments equipped with a 355 nm UV laser.

Filter: 560/40 or 585/15 
Relative brightness: Bright*

Sample data:

BD Horizon Brilliant™ Ultraviolet 661 (BUV661) (Ex Max 350 nm/Em Max 660 nm) is a tandem dye that combines BUV395 and an acceptor dye with an emission maximum at 660 nm. As part of the BD Horizon Brilliant™ Ultraviolet family, BUV661 provides an additional option for multicolor panels utilizing UV excitable dyes. The acceptor dye can be excited by the red (628–640 nm) laser resulting in cross-laser excitation and fluorescence spillover.

Filter: 670/25
Relative brightness: Bright*
Sample data:

BD Horizon Brilliant Violet™ 421 (BV421) (Ex Max 407 nm/Em Max 423 nm) is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BV421, Pacific Blue™ and BD Horizon™ V450 cannot be used simultaneously.

Filter: 431/28 or 450/50

Relative brightness: Very Bright*

Sample data:

BD Horizon Brilliant Violet™ 510 (BV510) (Ex Max 327 nm / 405 nm/Em Max 512 nm) is part of the BD Horizon Brilliant Violet™ family of dyes. This Sirigen base dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 327 nm / 405 nm and an emission maximum (Em Max) at 512 nm.  BD Horizon™ BV510, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 510 nm (e.g., a 525/50 bandpass filter). The dye can be excited by the UV (355 nm) laser resulting in cross-laser excitation and spillover. BD Horizon™ BV510 can be detected in the BD Horizon™ V500 (525/50 nm) filter set.

Filter: 525/50 
Relative brightness: Moderate*
Sample data:

BD Horizon Brilliant Violet™ 650 (BV650) (Ex Max 406 nm/Em Max 649 nm) is a tandem fluorochrome of BD Horizon™ BV421 and an acceptor dye with an Em Max at 650 nm. Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor™ 700 detectors. BD Horizon™ BV650 will have moderate spillover into the BD Horizon™ BV711 detector.

Filter: 660/20 
Relative brightness: Very Bright*

BD Horizon Brilliant Violet™ 750 (BV750) (Ex Max 409 nm/Em Max 754 nm) is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BD Horizon™ BV421 donor with an excitation maximum (Ex Max) of 409 nm and an acceptor dye with an emission maximum (Em Max) at 754 nm. BD Horizon™ BV750, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 750 nm (e.g., a 750/30 nm bandpass filter).

Filter: 750/30 
Relative brightness: Moderate*

*Relative resolution for each dye was determined based on the average stain index measured across multiple specificities and instruments. Relative resolution and absolute stain index values may change depending on instrument platform and instrument configuration, including lasers, laser power and filters.

BD Pharmingen™ FITC (FITC) (Ex Max 494 nm/Em Max 518 nm), fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor™ 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.

Filter: 530/30 
Relative brightness: Moderate*

BD Horizon Brilliant™ Blue 660-P2 (BB660-P2) (Ex Max 476 nm/Em Max 660 nm) is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a Sirigen polymer donor with an excitation maximum (Ex Max) of 476 nm and an acceptor dye with an emission maximum (Em Max) at 660 nm. BD Horizon™ BB630-P2, driven by BD innovation, is designed to be excited by the blue laser (488 nm) and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). The donor dye can be partially excited by the violet (405 nm) laser and the acceptor dye can be excited by the red laser (627–640 nm) resulting in cross-laser excitation and fluorescence spillover.

Filter: 660/20 
Relative brightness: Very Bright*

BD Horizon Brilliant™ Blue 755-P (BB755-P) (Ex Max 478 nm/Em Max 755 nm) is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a Sirigen polymer donor with an excitation maximum (Ex Max) of 478 nm and an acceptor dye with an emission maximum (Em Max) at 755 nm. BD Horizon™ BB755-P, driven by BD innovation, is designed to be excited by the blue laser (488 nm) and detected using an optical filter centered near 755 nm (e.g., a 750/50 nm bandpass filter). The donor dye can be partially excited by the violet (405 nm) laser and the acceptor dye can be excited by the red (627–640) laser resulting in cross-laser excitation and fluorescence spillover.

Filter: 750/50
Relative brightness: Bright*

*Relative resolution for each dye was determined based on the average stain index measured across multiple specificities and instruments. Relative resolution and absolute stain index values may change depending on instrument platform and instrument configuration, including lasers, laser power and filters.

BD Horizon Brilliant™ Yellow-Green 584 (BYG584) (Ex Max  563 nm/Em Max 583 nm) is part of the BD Horizon Brilliant™ Yellow-Green family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 563 nm and an emission maximum (Em Max) at 583 nm. BD Horizon™ BYG584, driven by BD innovation, is designed to be excited by the yellow-green laser (561 nm) and detected using an optical filter centered near 585 nm (e.g., 585/29 nm bandpass filter).

Filter: 585/29
Relative brightness: Very Bright*

BD Horizon™ PE-CF594 (PE-CF594) (Ex Max 496 nm and 566 nm/Em Max 615 nm) is a tandem conjugate that combines PE and CF594. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488 nm), green (532 nm) and yellow-green (561 nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20 nm bandpass filter). The donor dye can be excited by the blue (488 nm), green (532 nm) and yellow-green (561 nm) lasers and the acceptor dye can be excited by the green (532 nm) laser resulting in cross-laser excitation and fluorescence spillover. PE-CF594 is a brighter alternative to PE-Texas Red, with improved spectral characteristics. PE-CF594 reagents exhibit very consistent spillover values lot-to-lot, making them an ideal choice for the PE-Texas Red detector (610/20 nm).

Filter: 610/20 
Relative brightness: Very Bright*

Sample data:

BD Pharmingen™ PE-Cy7 (PE-Cy7) (Ex Max 496 nm and 566 nm/Em Max 781 nm) is a tandem fluorochrome that combines PE and a cyanine dye. PE can be excited by the blue (488 nm), green (532 nm) and yellow-green (561 nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60 nm bandpass filter). The donor dye can be excited by the blue (488 nm), green (532 nm) and yellow-green (561 nm) lasers and the acceptor dye can be excited by the red (627-640 nm) laser resulting in cross-laser excitation and fluorescence spillover. PE-Cy7 conjugated reagents are as bright as PE conjugates. PE-Cy7 is particularly sensitive to photo-induced degradation, resulting in loss of fluorescence and changes in fluorescence spillover. Extreme caution must be taken to avoid light exposure and prolonged exposure to paraformaldehyde fixative. Fixed cells should be analyzed within 4 hours of fixation in paraformaldehyde or transferred to a paraformaldehyde-free buffer for overnight storage.

Filter: 760/60 
Relative brightness: Very Bright*

Allophycocyanin (APC) (Ex Max 651 nm/Em Max 660 nm) is an accessory photosynthetic pigment found in blue-green algae. Its molecular weight is approximately 105 kDa. APC has six phycocyanobilin chromophores per molecule, which make it a very bright fluorochrome that is highly suitable for flow cytometry applications. APC is designed to be excited by the red (627–640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor™ 647 cannot be used simultaneously.

Filter: 660/20 
Relative brightness: Bright*

BD Horizon™ Red 718 (R718) (Ex Max 695 nm/Em Max 718 nm) is part of the BD Horizon™ Red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 695 nm and an emission maximum (Em Max) at 718 nm. R718, driven by BD innovation, is designed to be excited by the red laser (627–640 nm) and detected using an optical filter centered near 720 nm (e.g., a 720/40 nm bandpass filter).

Filter: 720/40

Relative brightness: Bright*

Sample data: