BD FACSCelesta™ Flow Cytometer Configuration Sheet: Blue ...
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BD FACSCelesta ™ Flow Cytometer Configuration Sheet Blue-Violet-Red (BVR) Laser Configuration The BD FACSCelesta™ flow cytometer is designed to simplify the use of multicolor flow cytometry and allow researchers to benefit from new innovations in instrument and reagent technology. This platform offers multiple configurations to meet varied application needs. The BVR configuration of the BD FACSCelesta includes blue, violet, and red lasers, optimized to get the most out of the legacy fluorochromes, viability dyes, and BD Horizon Brilliant™ dyes shown in the optical path diagrams. While a blue laser has long been standard in flow cytometry, the violet laser is gaining popularity as more bright violet-excited fluorochromes, such as BD Horizon Brilliant™ Violet reagents, are introduced and enthusiastically adopted. The red laser, which optimally excites allophycocyanin (APC) and APC-based tandem dyes, allows markers to be spread across more lasers, further reducing the overall compensation needed for a multicolor panel. Multicolor flow cytometry panel design has presented challenges for researchers, such as varying marker expression, varying dye brightness, and significant emission spillover between fluorescence channels. The combination of bright, narrow-spectrum, BD Horizon Brilliant fluorochromes and sensitive optics results in panels that can readily resolve even dim populations, yet are easy to use and compensate. The system operates with BD FACSDiva™ software, a collection of convenient and easy-to-use tools for flow cytometer and application setup, data acquisition, and data analysis. Blue laser Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 520, 570, 620 Violet laser Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 450, 510 Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 660, 700, 780 Red laser Optical path diagrams and fluorochrome support Laser polygons show fluorochromes, mirrors, filters, and optical paths for the BD FACSCelesta BVR configuration. PE-CF594 PI 610/20 BP SSC BB515 FITC Alexa Fluor® 488 530/30 BP PerCP PerCP-Cy™5.5 7-AAD 695/40 BP PE 575/26 BP APC-Cy ™ 7 780/60 BP APC 670/30 BP APC-R700 730/45 BP BV786 780/60 BP BV605 610/20 BP BV650 660/20 BP BV421 V450 Pacific Blue™ 450/40 BP BV510 V500 525/50 BP
Transcript of BD FACSCelesta™ Flow Cytometer Configuration Sheet: Blue ...
BD FACSCelesta™ Flow Cytometer Configuration Sheet Blue-Violet-Red (BVR) Laser Configuration
The BD FACSCelesta™ flow cytometer is designed to simplify the use of multicolor flow cytometry and allow researchers to benefit from new innovations in instrument and reagent technology. This platform offers multiple configurations to meet varied application needs.
The BVR configuration of the BD FACSCelesta includes blue, violet, and red lasers, optimized to get the most out of the legacy fluorochromes, viability dyes, and BD Horizon Brilliant™ dyes shown in the optical path diagrams. While a blue laser has long been standard in flow cytometry, the violet laser is gaining popularity as more bright violet-excited fluorochromes, such as BD Horizon Brilliant™ Violet reagents, are introduced and enthusiastically adopted. The red laser, which optimally excites allophycocyanin (APC) and APC-based tandem dyes, allows markers to be spread across more lasers, further reducing the overall compensation needed for a multicolor panel.
Multicolor flow cytometry panel design has presented challenges for researchers, such as varying marker expression, varying dye brightness, and significant emission
spillover between fluorescence channels. The combination of bright, narrow-spectrum, BD Horizon Brilliant fluorochromes and sensitive optics results in panels that can readily resolve even dim populations, yet are easy to use and compensate. The system operates with BD FACSDiva™ software, a collection of convenient and easy-to-use tools for flow cytometer and application setup, data acquisition, and data analysis.
Blue laser
Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 520, 570, 620
Violet laser
Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 450, 510
Other fluorochromes supported: BD Horizon™ Fixable Viability Stain 660, 700, 780
Red laser
Optical path diagrams and fluorochrome supportLaser polygons show fluorochromes, mirrors, filters, and optical paths for the BD FACSCelesta BVR configuration.
PE-CF594PI
610/20 BP
SSC
BB515FITC
Alexa Fluor® 488530/30 BP
PerCPPerCP-Cy™5.5
7-AAD695/40 BP
PE575/26 BP
APC-Cy™7780/60 BP
APC670/30 BP
APC-R700730/45 BP
BV786780/60 BP
BV605610/20 BP
BV650660/20 BP
BV421V450
Pacific Blue™450/40 BP
BV510V500
525/50 BP
BD FACSCelesta Configuration Sheet Blue-Violet-Red (BVR) Laser Configuration
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10-color T-cell analysis on the BD FACSCelesta BVR configuration
This T-cell panel demonstrates the sensitivity and resolution of the BD FACSCelesta system, even in detecting rare subpopulations. After normal human whole blood was washed and lysed, BD Horizon Brilliant and traditional dyes were used to identify rare T-cell and Treg subpopulations. A. Cells were gated to select the CD3+ T cells. B. CD3+ lymphocytes were gated to show the CD4+ helper T cells and CD8+ cytotoxic T cells. C. Gated on the CD4+ T cells, surface markers were used to identify CD25+CD127– Tregs. D. Gated on Tregs, surface markers were used to identify memory and naïve Treg subsets. E. CD4+ helper T cells were analyzed for memory T-cell subsets using CD45RO and CD197 (Left) and Effector Memory subsets were further defined with surface markers for CD27 and CD194 (Right). F. CD8+ cytotoxic T cells were analyzed for Effector and memory subsets using CD45RO and CD197 (Left) and Effector/effector memory subsets were further defined using CD27 and CD28 surface markers. Similar analysis was completed for CD8+ cytotoxic T cells (Right).
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310
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102 103 104 105-98
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CD197 BV521
CD45
RO B
B515
Effector Memory CD4 Cells
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CD28 BV605
CD27
APC
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0 102 103 104 105-139
-304
CD45RO BB515
CD19
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Memory Tregs
NaïveTregs
Effector/Effector Memory CD6 Cells
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CD197 BV421
CD45
RO B
B515
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CD194 PE
CD27
APC
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CD25 PE-CF594
CD12
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CD3 APC-Cy7
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CD4 BV510
CD8
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5.5 CD8+ T Cells
CD4+ T Cells
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B
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BD FACSCelesta Configuration Sheet Blue-Violet-Red (BVR) Laser Configuration
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Two-color flow cytometric analysis of apoptosis and viability in Jurkat cells
Jurkat cells were treated with 0.025% DMSO vehicle (top plots) or 5 μM camptothecin (bottom plots) for 4 hours, harvested from culture, washed, and resuspended in Annexin V Binding Buffer. Cells were then incubated with BD Pharmingen™ APC Annexin V (Cat. No. 550475) and BD Pharmingen™ Propidium Iodide Staining Solution (Cat. No. 556463) (PI) for 15 minutes at room temperature protected from light, and then analyzed by flow cytometry on a BD FACSCelesta system. Debris was excluded based on the light scatter properties of Jurkat cells (left plots). DMSO vehicle-treated cells were primarily Annexin V–PI–, indicating that most cells were live. Camptothecin-treated cells show an increase in the number of Annexin V+ (apoptotic) and Annexin V+PI+ (dead) cells, indicating that camptothecin treatment induced apoptosis and cell death.
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Annexin V APC
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idiu
m Io
dide
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Live
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015
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FSC
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(x1,000)
(x1,
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CellsD
MSO
Cam
ptot
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102-102 103 104 105-472
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Annexin V APC
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idiu
m Io
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Dead
Apoptosis
Live
50
5010
015
020
025
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100 150 200 250
FSC
SSC
(x1,000)
(x1,
000)
Cells
BD FACSCelesta Configuration Sheet Blue-Violet-Red (BVR) Laser Configuration
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.BD flow cytometers are Class 1 Laser Products.Alexa Fluor® is a registered trademark and Pacific Blue™ is a trademark of Life Technologies Corporation.CF™ is a trademark of Biotium, Inc.Cy™ is a trademark of GE Healthcare. Cy™ dyes are subject to proprietary rights of GE Healthcare and Carnegie Mellon University, and are made and sold under license from GE Healthcare only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from GE Healthcare, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2015 BD23-17600-00
Description Cat. No.
BD FACSCelesta™ Flow Cytometer, BVR Configuration 660344
BD FACSCelesta™ High Throughput Sampler (HTS) Option 658946
BD FACSFlow™ Supply System 649908
BD FACSCelesta™ Standard Workstation Bundle 660472
Excitation Laser Fluorochrome Exmax Emmax Relative Brightness
Violet (405 nm)
BD Horizon Brilliant™ Violet 786 (BV786) 407 nm 786 nm
BD Horizon Brilliant™ Violet 650 (BV650) 407 nm 650 nm
BD Horizon Brilliant™ Violet 605 (BV605) 407 nm 602 nm
BD Horizon Brilliant™ Violet 510 (BV510) 405 nm 510 nm
BD Horizon Brilliant™ Violet 421 (BV421) 407 nm 421 nm
Blue (488 nm)
PerCP 482 nm 678 nm
PerCP-Cy™5.5 482 nm 695 nm
BD Horizon™ PE-CF594 496 nm 612 nm
PE 496 nm 578 nm
BD Horizon Brilliant™ Blue 515 (BB515) 490 nm 515 nm
FITC 494 nm 520 nm
Alexa Fluor® 488 495 nm 519 nm
Red (640 nm)
APC-Cy™7 650 nm 785 nm
APC-R700 652 nm 704 nm
APC 660 nm 660 nm
Ordering Information
Relative Brightness Key: Dim
Moderate
Bright
Brightest
