BD Biosciences Fluorochrome Reference Chart

Visit bdbiosciences.com/colors for detailed information about our newest fluorochromes and instrumentation.
To select your optimal combination of fluorochromes, visit bdbiosciences.com/spectra to use an interactive fluorescence spectrum tool.
23-9582-08

Instrument
Excitation Laser Line
(nm) Fluorescence Channel Fluorochromes provided by BD Biosciences
Brightness of various fluorochrome conjugates
BD Accuri™ C6 488 FL1 Green FITC Alexa Fluor® 488
FL2 Yellow PE PI Relative
FL3 Red 7-AAD PerCP PerCP-Cy™5.5 PE-Cy™7 Brightness Reagent Filter
640 FL4 Red APC Alexa Fluor® 647
BD FACSCalibur™ 488 FL1 Green FITC Alexa Fluor® 488 Brilliant Violet™ 421 450/50
FL2 Yellow PE PI
FL3 Red 7-AAD PE-Cy™5 PerCP PerCP-Cy5.5 PE-Cy7
635 FL4 Red APC Alexa Fluor® 647
PE 575/26
BD FACSVerse™* 488 Green FITC Alexa Fluor® 488
Brilliant Violet 605 610/20

BRIGHTEST
Yellow PE PI
Orange BD Horizon™ PE-CF594a PE-Texas Red®a
Red 7-AAD PE-Cy5 PerCP PerCP-Cy5.5
Infrared PE-Cy7 BD Horizon PE-CF594 610/20
640a Red APC Alexa Fluor® 647
Far Red Alexa Fluor® 700a
PE-Cy5 670/14
Infrared BD APC-H7 APC-Cy7
405a Blue Brilliant Violet™ 421 BD Horizon™ V450 VPD450 Pacific Blue™
Green BD Horizon™ V500 AmCyan APC 660/20
BD FACSCanto™ II 488 Green FITC Alexa Fluor® 488
Yellow PE PI
Orange BD Horizon PE-CF594a PE-Texas Red®a
PE-Cy7 780/60
Red 7-AAD PE-Cy5 PerCP PerCP-Cy5.5

BRIGHT
Infrared PE-Cy7 Alexa Fluor® 647 660/20
561b Yellow PE PI
Orange BD Horizon PE-CF594 PE-Texas Red®
Red PE-Cy5 PerCP-Cy5.5 695/40
Infrared PE-Cy7
633 Red APC Alexa Fluor® 647
Alexa Fluor® 488 530/30
Far Red Alexa Fluor® 700a
Infrared BD APC-H7 APC-Cy7
FITC 530/30

MODERATE
405 a
Blue Brilliant Violet 421
™
BD Horizon™ V450 VPD450 Pacific Blue™
Green BD Horizon V500 BD Horizon V500-C AmCyan
BD LSRFortessa™ and 488 Green FITC Alexa Fluor® 488
Special Order Yellow PE PI
BD Horizon V450 450/50
BD LSRFortessa Orange BD Horizon PE-CF594 PE-Texas Red®
(typical setup)b Red 7-AAD PE-Cy5 PerCP PerCP-Cy5.5 Pacific Blue™ 450/50
Infrared PE-Cy7
532b or 561b Yellow PE PI
Orange BD Horizon PE-CF594 PE-Texas Red® Alexa Fluor® 700 730/45
Red PE-Cy5
Infrared PE-Cy7
PerCP 695/40
640 Red APC Alexa Fluor® 647
Far Red Alexa Fluor® 700
Infrared BD APC-H7 APC-Cy7 APC-Cy7 780/60

DIM
405 Blue Brilliant Violet 421
™
BD Horizon V450
™
VPD450 Pacific Blue ™

Green BD Horizon V500 AmCyan

Orange Brilliant Violet™ 605a
AmCyan 525/20
355 Blue Hoechst 33342
BD FACSAria™ III and 488 Green FITC Alexa Fluor® 488 BD Horizon V500 525/20
Special Order BD FACSAria Yellow PE PI
(typical setup)b Orange BD Horizon PE-CF594 PE-Texas Red®
Red 7-AAD PE-Cy5 PerCP PerCP-Cy5.5 BD APC-H7 780/60
Infrared PE-Cy7
561 Yellow PE PI Freshly isolated lymphocytes, stained with anti-human CD4 (RPA-T4) conjugated with various
Orange BD Horizon PE-CF594 PE-Texas Red® fluorochromes run on a BD LSR II flow cytometer. The fluorochromes were ranked based on
Red PE-Cy5 observed stain index values. This chart is meant as a guideline of relative stain indices of various
fluorochromes. Observed relative stain indices may vary depending on instrument, instrument
Infrared PE-Cy7
configuration, reagents, and cell type used.
640 Red APC Alexa Fluor® 647
Far Red Alexa Fluor® 700
Infrared BD APC-H7 APC-Cy7
405 Blue Brilliant Violet™ 421 BD Horizon™ V450 VPD450 Pacific Blue™
Green BD Horizon V500 AmCyan
Orange Brilliant Violet 605
™ a

375b Blue Hoechst 33342

BD Influx™ 488 Green
Yellow
FITC
PE
Alexa Fluor® 488
PI
W
Orange BD Horizon PE-CF594 PE-Texas Red®
Red 7-AAD PE-Cy5 PerCP PerCP-Cy5.5
Infrared PE-Cy7
532 or 561 Yellow PE PI
Orange BD Horizon PE-CF594 PE-Texas Red®
Red PE-Cy5
Infrared PE-Cy7
640 Red APC Alexa Fluor® 647
Far Red Alexa Fluor®700

405
Infrared
Blue
BD APC-H7
Brilliant Violet 421
™
APC-Cy7
BD Horizon™ V450 VPD450 Pacific Blue™
D
Green BD Horizon V500 AmCyan
Orange Brilliant Violet™ 605a
375 Blue Hoechst 33342
BD FACSJazz™ 488 Green FITC Alexa Fluor® 488
Yellow PE
Red PerCP-Cy5.5
Infrared PE-Cy7 Stain Index = D/W
640 a
Red APC Alexa Fluor® 647 Resolution sensitivity (the ability to resolve a dim positive signal from background) is a function
of the difference between positive and background peak means (D) and the spread of the
Infrared BD APC-H7 APC-Cy7
background peak (W). The stain index is a metric that captures both of these factors.
405 a
Blue Brilliant Violet 421
™
BD Horizon™ V450 Pacific Blue™
Green BD Horizon V500
a
Available through laser and/or detector options.
b
More laser and detector options are available through the Special Order Research Products (SORP) program.

Choose a winning combination - Guidelines for selecting reagents for multicolor flow cytometry
1 The basics: Know your

instrument
Reagent selection starts with your
2 Fluorochromes:
Go for the bright
Rank available dyes according to their
3 Minimize spillover
As soon as cells are stained with
multiple reagents, spectral overlap
4 Colors and specificities:
Define winning combinations
Once the fluorochromes to be used
5 Tandem dyes
APC-Cy7, and to a lesser extent,
PE-Cy7, can degrade in the presence of
6 Validation
Use controls (such as fluorescence-
minus-one, or FMO) to validate
instrument configuration. The lasers and intrinsic brightness on a particular (or spillover) becomes an issue. The have been defined, you can begin to light, fixative, and elevated temperatures your selected multicolor reagent
detectors in your configuration dictate instrument (when configured with a more colors you attempt to resolve match antibody specificities to particular so that they emit in the parent dye cocktail. FMO controls help define
how well your cytometer can excite and specified set of lasers and filters). on any particular cell, the more fluorochromes. Generally, reserve detector (APC or PE). By minimizing the contribution of spillover to the
measure a given fluorochrome, and spillover impacts sensitivity. We use the brightest fluorochromes for dim the exposure of samples to light, heat, background in a given detector, and
whether you have enough detectors compensation, an adjustment applied antigens, and vice versa, but avoid and formaldehyde-based fixatives, this are therefore useful in gauging the
to read out a given combination of to all colors, to correct for spillover. spillover from bright cell populations problem can be largely avoided. For sensitivity of that detector in the context
fluorochromes. For example, a cell population into detectors requiring high sensitivity more stable tandem dyes, BD now offers of a certain reagent cocktail.
fluorescing only in FITC will show no PE for those populations. BD APC-H7 conjugated antibodies.
fluorescence, on average, but will likely
exhibit more spread in the PE detector
after compensation than completely For additional guidelines, visit bdbiosciences.com/colors to download the
unstained cells. Application Note “Selecting Reagents for Multicolor Flow Cytometry.”

* Capable of detecting 8 colors simultaneously (4 blue laser, 2 red laser, 2 violet laser)
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
APC-Cy7: US patent 5,714,386
Brilliant Violet™ is a trademark of Sirigen Group Ltd.
Cy™ is a trademark of Amersham Biosciences Corp. Cy dyes are subject to proprietary rights of Amersham Biosciences Corp and Carnegie Mellon University and are made and sold under license from Amersham
Biosciences Corp only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.
Pacific Blue™ is a trademark, and Alexa Fluor® and Texas Red® are registered trademarks of Molecular Probes, Inc.
CF™ is a trademark of Biotium, Inc.
BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2012 BD