CytoFLEX Family Brochure

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1 CytoFLEX Family

2 CytoFLEX CytoFLEX CytoFLEX CytoFLEX CytoFLEX 1 CytoFLEX S 1 CytoFLEX LX 21 2 EVERY ERY Event Matters ters

3 CytoFLEX 1 T T IR CD8 IR CytoFLEX / Karen Hogg, PhD, Bioscience Technology Facility, University of York

CytoFLEX Wavelength Division Multiplexer WDM WDM WDM

detectors APD 1 8 nm8 CytoFLEX IC CytoFLEX WDM APD IC

One (FMO) CytoFLEX CytoFLEX FITC PE PECy7 APC Ryan

4 CytoFLEX Wavelength Division Multiplexer WDM WDM WDM Photomultiplier tubes PMT Avalanche Photodiode detectors APD 1 8 nm8 CytoFLEX IC CytoFLEX WDM APD IC Human PBMCs Mouse Spleen T T FOXP+Fluorescence Minus One (FMO) CytoFLEX CytoFLEX FITC PE PECy7 APC Ryan Duggan, UC Flow Core Lab Director FITC PE PC7 APC PB EVERY Event Matters

CytoFLEX A B C D E F G H Microparticle MP Megamix-Plus FSC &

Gigamix Gigamix SMP Submicron particles 7 9 nm nm nm 2 nm 2 nm 1 nm

nmssc nmvssc 2 Large MP..22 m MP..22 m VSSC 2 S..1 m Max.

C,9 F, VSSC S Max G 1, H 1,22 CytoFLEX BioCytex Stago Group

Submicron particle analysis and counting is highly favored by the

cytometer CytoFLEX 1 nm * 1 Andreas Spittler, MD, Associate

Facility Flow Cytometry & Department of Surgery, Research

5 CytoFLEX A B C D E F G H Microparticle MP Megamix-Plus FSC & Megamix-Plus SSC BioCytex Refs # 782 and # 78 respectively V/V Gigamix Gigamix SMP Submicron particles 7 9 nm nm nm 2 nm 2 nm 1 nm nm 7 nm Gigamix A 88 nm SSC B nm VSSC 88 nm SSC nm VSSC BxPC 88 nmssc nmvssc 2 Large MP..22 m MP..22 m VSSC 2 S..1 m Max..8 m 7 nm nm Annexin-V-FITC SBTF1-PE CD nm SSC nm VSSC MP C,9 F, VSSC S Max G 1, H 1,22 CytoFLEX BioCytex Stago Group Philippe Poncelet Aix-Marseille VRCM Stéphane Robert CYTO 21 Submicron particle analysis and counting is highly favored by the use of side scatter from the violet laser on the CytoFLEX flow cytometer CytoFLEX 1 nm * 1 Andreas Spittler, MD, Associate Professor for Pathophysiology, Medical University of Vienna, Core Facility Flow Cytometry & Department of Surgery, Research Laboratories *2nm Set-Up of the CytoFLEX for Extracellular Vesicle Measurement Andreas Spittler Violet SSC-H [A] FSC-H / Violet SSC-H FSC-H

6 CytoFLEX APD Avalanche Photodiode detector APD 1 2 CytoFLEX minimum R-squared value.9998 Loïc Tauzin, Valerie Glutz and Miguel Garcia, Ecole Polytechnique Federale De Lausanne Flow Cytometry Core Facility EVERY Event Matters

7 CytExpert CytExpert 21 CFR Part11 CytExpert QC QC D 1 1 CPU OS USB Core i ( MB Cache, 2.9 GHz) Windows 7, 8, Professional, bit GB RAM GB / GB 1,92 1,8 USB 2. USB / 9 2 U V 9 7

8 CytoFLEX CytoFLEX Blue-Red-Violet B-R-V 88 nm Blue 8 nm Red nm Violet 1 / 8/2 / (2) 712/2 2/ (2) /2 (2) 9/ 78/ () Blue-Red-Violet B-R-V 88 nm (Blue) 8 nm (Red) nm (Violet) B 1 B1 12 B2 12 B 11 B 11 B B 2 B7 2 B7 9 B8 9 2 B B9 8 2 C B B11 2 B1 2 2 B12 2 C C B18 1 B1 2 2 B19 1 B1 2 1 B B Blue 88nm Red 8nm Rainbow FITC-A PE-A ECD-A PC.-A PC7-A APC-A APC- 8-speak SPHERO TM Rainbow Calibration Particles 8 EVERY Event Matters

Lab Manager North Carolina State Veterinary College Violet

9 Violet Side Scatter CytoFLEX 88 nm nm nm 88 nm nm Violet Side Scatter VSSC nm CytoFLEX 2 Violet Sarah Schuett Core Lab Manager North Carolina State Veterinary College Violet nm P1 2 A7-A APC-A7-A 2 PB-A 2 KO2-A Violet-A Violet-A Violet78-A 9

10 CytoFLEX S CytoFLEX S Blue-Red-Violet-Yellow Green B-R-V-Y 88 nm Blue 2 8 nm Red nm Violet 1 nm Yellow Green 1 / 8/2 / (2) 712/2 2/ (2) /2 (2) 9/ (2) 78/ (2) Blue-Red-Violet-Yellow Green B-R-V-Y 88 nm (Blue) 8 nm (Red) nm (Violet) 1 nm (Yellow Green) B B92 2 B B C B C nm Yellow Green RFP RFP DsRed HcRed 88 nm GFP YFP DsRed HcRed DsRed HcRed DsRed YFP YFP GFP Yellow Green 1nm PE-A ECD-A PC.-A PC7-A 8-speak SPHERO TM Rainbow Calibration Particles EVERY Event Matters

Blue-Red-Violet-Near UV B-R-V-N 88 nm Blue 8 nm Red nm Violet 7 nm Near UV

B-R-V-N 88 nm (Blue) 8 nm (Red) nm (Violet) 7 nm (Near UV) B787 1 2 B789 2

Cycle Violet nm FITC PE 7 nm Hoechst Side population 7 nm Side population

11 Blue-Red-Violet-Near UV B-R-V-N 88 nm Blue 8 nm Red nm Violet 7 nm Near UV 2 1 / (2) 8/2 / 9/ 78/ (2) 2/ (2) /2 (2) 7/ 712/2 Blue-Red-Violet-Near UV B-R-V-N 88 nm (Blue) 8 nm (Red) nm (Violet) 7 nm (Near UV) B B789 2 B UV 7 nm Near UV Hoechst DAPI Brilliant UV Side population Dye Cycle Violet nm FITC PE 7 nm Hoechst Side population 7 nm Side population Near UV 7nm DAPI-A HoechstRed-A 8-speak SPHERO TM Rainbow Calibration Particles 11

8 nm (Red) nm (Violet) 88 nm (Infrared) C111 1 2 C11 2 C119 9 2 C118 2 2 Infrared 88 nm

12 CytoFLEX S Blue-Red-Violet-Infrared B-R-V-I 88 nm Blue 8 nm Red nm Violet 88 nm Infrared ) 2 1 / 8/2 / (2) 712/2 8/2 2/ (2) /2 9/ 7/ (2) 88/ Blue-Red-Violet-Infrared B-R-V-I 88 nm (Blue) 8 nm (Red) nm (Violet) 88 nm (Infrared) C C11 2 C C Infrared 88 nm Infrared 88nm SPHERO TM Fluorescent IR Flow Cytometer Particles AF79-A PF8-A 12 EVERY Event Matters

Blue-Violet-Yellow Green-Near UV B-V-Y-N 88 nm Blue 2 nm

(Yellow Green) 7 nm (Near UV) B78 12 2 2 B919 2 B781 8 2

13 Blue-Violet-Yellow Green-Near UV B-V-Y-N 88 nm Blue 2 nm Violet 1 nm Yellow Green ) 7 nm Near UV ) 2 12 / (2) 8/2 / 9/ (2) 2/ (2) /2 (2) 7/ 78/ Blue-Violet-Yellow Green-Near UV B-V-Y-N 88 nm (Blue) nm (Violet) 1 nm (Yellow Green) 7 nm (Near UV) B B919 2 B B C Biomek i-series CytoFLEX Biomek CytoFLEX ID CytoFLEX 1

14 CytoFLEX LX CytoFLEX LX 21 Blue-Red-Violet-Yellow Green-Near UV-Infrared (B-R-V-Y-N-I) 88 nm Blue 8 nm Red nm Violet 1 nm Yellow Green ) 7 nm Near UV 88 nm Infrared 2 21 / / (2) 2/ () 8/2 /2 () / (2) 7/ (2) 9/ 7/ 712/2 7/ () 8/2 88/ Blue-Red-Violet-Yellow Green-Near UV-Infrared B-R-V-Y-N-I 88 nm (Blue) 8 nm (Red) nm (Violet) 1 nm (Yellow Green) 7 nm (Near UV) 88 nm (Infrared) C 21 2 C 19 A 8 FTTC / Traditional filter used for APC detection on 8 nm APC PE PE-Cy7 PE-Texas Red PE-Cy (88 nm excitation) PE-Cy (8 nm excitation) Wavelength (nm) B 2/ Pacific Blue / PE-Cy7 Pacific Blue PE-Texas Red BUV9 / 2 PE-Texas Red APC PE-Cy PE-Alexa Fluor 79 2 BUV9 PE-Texas Red APC PE-Cy PE-Alexa Fluor 79 2 Pacific Blue BUV9 APC PE-Alexa Fluor Wavelength (nm) Wavelength (nm) Wavelength (nm) PE-Texas Red 7/ PE-Cy7 / 8/2 8 8 APC PE-Cy PE-Alexa Fluor 79 2 Pacific Blue BUV9 APC PE-Alexa Fluor 79 Pacific Blue BUV9 PE-Texas Red Wavelength (nm) Wavelength (nm) Wavelength (nm) A 88 nm 8 nm FITC PE PE-Texas Red APC, PE-Cy APC-Cy7 2 PE-Cy APC B CytoFLEX LX BUV9 Pacific Blue PE-Texas Red PE-Cy7 APC AF79 1 EVERY Event Matters

Blue-Red-Violet-Yellow-UV-Infrared B-R-V-Y-U-I 88 nm Blue 8 nm Red nm Violet 1 nm Yellow Green ) nm UV

Violet SSC-H Brilliant UV UV NUV Versa Lyse CytoFLEX LX BUV9 BUV9 donor 1 BUV1 donor 2 ) Lymphos ( )

15 Blue-Red-Violet-Yellow-UV-Infrared B-R-V-Y-U-I 88 nm Blue 8 nm Red nm Violet 1 nm Yellow Green ) nm UV 88 nm Infrared 2 21 / / / 2/ () 8/2 /2 () / (2) 7/ (2) 9/ 7/ 7/ () 712/2 7/ () 8/2 88/ Blue-Red-Violet-Yellow-UV-Infrared B-R-V-Y-U-I 88 NM (Blue) 8 NM (Red) NM (Violet) 1 NM (Yellow Green) NM (UV) 88 NM (Infrared) C C C C UV nm CD-BUV9 CD-BUV9 CD-BUV1 [A] FSC-H / Violet SSC-H Brilliant UV UV NUV Versa Lyse CytoFLEX LX BUV9 BUV9 donor 1 BUV1 donor 2 ) Lymphos ( ) CD-BUV9-UV-A CD-BUV9-UV2-A CD-BUV1-UV7-A NUV 7 nm CD-BUV9-NUV-A CD-BUV9-NUV2-A CD-BUV1-NUV7-A UV nm UV-A UV2-A UV7-A 1

16 / / / B7129 CytoFLEX, 11 mm B2 CytoFLEX Daily QC Fluorospheres A-1- CytoFLEX, 11 mm C17 CytoFLEX Daily IR QC Fluorospheres A-1-8 Deep Clean Solution Bottle Kits B1 CytoFLEX Sheath Fluid A-1- Sheath Bottle Kit A9 FlowClean A-1-7 Waste Bottle Kit 98 9 PS 771 WASTE CONTAINER, L PP V B89 L Sheath/Waste Line Kit B21 Plate Loader Sample Probe (with tubing to attach to plate assembly) A-1-8 CytoFLEX B9289 CytoFLEX Installation Kit A-1-1 CytoFLEX A1-1-8 / nm Bandpass Filter B7117 9/2 nm Bandpass Filter B991 / nm Bandpass Filter A1-1- /2 nm Bandpass Filter A1-1-9 / nm Bandpass Filter B9297 /2 nm Bandpass with OD1 Filter B9 / nm Bandpass with OD1 Filter A1-1- 8/ nm Bandpass Filter A /8 nm Bandpass Filter A1-1- / nm Bandpass Filter B7128 /2 nm Bandpass Filter B782 7/ nm Bandpass Filter B929 /2 nm Bandpass with OD1 Filter A1-1- 9/ nm Bandpass Filter B712 1/2 nm Bandpass Filter B792 7/ nm Bandpass Filter A / nm Bandpass Filter A /2 nm Bandpass Filter B9 2/ nm Bandpass with OD1 Filter B / nm Bandpass Filter B719 / nm Bandpass Filter B991 7/ nm Bandpass Filter B7227 1/ nm Bandpass Filter A / nm Bandpass Filter B7121 8/1 nm Bandpass Filter B / nm Bandpass Filter B789 8/ nm Bandpass Filter B991 8/2 nm Bandpass Filter A /2 nm Bandpass Filter B991 88/ nm Bandpass Filter 1 EVERY Event Matters

17 DuraClone Powered by 2 nm 88 nm 8 nm / 2/ 2/ 8/2 /2 9/ 78/ / 712/2 78/ PB KrO FITC PE ECD PC. PC7 APC AF7 AF7 DuraClone Immunophenotyping (IM) Basic Tube B9 APC- A7 APC- A7 - CD CD1 CD CD19 - CD1 CD - CD8 - CD - B Cell Tube B18 IgM CD IgD CD21 CD19 - CD27 CD CD8 - T Cell Subsets Tube B28 CD7 CD CDRA CCR7 CD28 PD1 CD27 CD - CD8 - CD - Dendritic Cells Tube B1 HLA-DR CD CD1 Lin** - CD1c CD11c Clec9A - - CD TCRs Tube B TCR Vδ2 CD TCR γδ TCR αβ HLA-DR - TCRVδ1 CD - CD8 - CD - Treg Tube B Helios CD CDRA CD2 - CD9 CD - FoxP - - CD - Granulocytes Tube B881 CD1 CD CD29 - CD1 CD CD11b PD-L1 - - Lin*** CD2L - Tube C12 AF7 - - CD ing Beads - 7-AAD DuraClone Immune Function (IF) T Activation B889 CD - IFN γ TNF α - - IL CD8 - - CD T Helper Cell C IL-17A - IFN γ IL- CD CD DuraClone Rare Event (RE) CLB Tube B89 CD2 CD CD81 ROR-1 - CD79b CD19 CD CD - PC Tube B89 CD8 CD CD81 CD27 - CD19 CD2 CD CD - ALB Tube C1 - CD CD8 - CD CD CD CD8 CD2 - ** CD/CD1/CD19/CD2/CD *** CD/1/CD19/CD 17

18 CytoFLEX CytoFLEX UV nm B-R-V-Y-U-I / 2/ 7/ BUV9 BUV9 Qdot 2 Qdot BUV1 Hoechst Red Near UV 7 nm B-V-Y-N B-R-V-N B-R-V-Y-N-I / 2/ 7/ Cascade Blue DAPI ebfp Hoechst Blue Marina Blue BUV9 BUV1 Hoechst Red Violet nm B-R-V B-R-V-Y B-R-V-N B-R-V-I B-V-Y-N B-R-V-Y-N-I B-R-V-Y-U-I / 2/ /2 / / 7/ 78/ Alexa Fluor V BV21 CFP Ctyophase Violet efluor Pacific Blue Zombie Violet AmCyan V BV Cascade Yellow ecfp Krome Orange LIVE/DEAD Fixable Aqua Zombie Aqua BV efluor 2NC Qdot BV Qdot BV78 BV78 Qdot 8 Blue 88 nm B-R-V 8/2 /2 9/ 78/ B-R-V-Y 9/ B-R-V-N /2 9/ 78/ 8/2 B-R-V-I 2/ 9/ 7/ B-V-Y-N 9/ B-R-V-Y-N-I /2 7/ 7/ B-R-V-Y-U-I 7/ Alexa Fluor 88 BB1 CFSE FITC Fluo- MitoSpy Green FM Oregon Green 88 Zombie Green Alexa Fluor Phycoerythrin (PE) Propidium iodide (PI) 7-AAD Alexa Fluor -R-PE PE-CF9 PE-Texas Red Propidium iodide (PI) Alexa Fluor 7-R-PE DRAQ7 PE/Cy PE/Cy. PerCP PerCP-Cy. Propidium iodide (PI) DRAQ7 PE/Cy7 Yellow Green 1 nm B-R-V-Y B-V-Y-N B-R-V-Y-N-I B-R-V-Y-U-I 8/2 /2 7/ 9/ 7/ 7/ 78/ Alexa Fluor Cy DsRed dtomato MitoSpy Orange CMTMRos Phycoerythrin (PE) Propidium iodide (PI) mcherry PE/Dazzle 9 PE-CF9 PE-Texas Red PHrodo Red Propidium iodide (PI) Zombie Red mplum PE/Cy PE-Alexa Fluor 8 PE/Cy PE/Cy. PerCP PerCP/Cy. Propidium iodide (PI) PE/Cy7 B-R-V B-R-V-Y 78/ Red 8 nm B-R-V-N B-R-V-I / 712/2 7/ B-R-V-Y-N-I B-R-V-Y-U-I 7/ Alexa Fluor Alexa Fluor Alexa Fluor 7 Allophycocyanin (APC) Cy efluor Flash Phalloidin NIR 7 Helix NP NIR TO-PRO- Alexa Fluor 8 Alexa Fluor 8-APC Alexa Fluor 8-R-PE Alexa Fluor 7-APC Alexa Fluor 7 Cy. Cy.-APC DRAQ DRAQ7 Alexa Fluor 7-APC APC- efluor 78 APC/Fire 7 APC-Cy7 APC-H7 DRAQ7 Zombie NIR Infrared 88 nm B-R-V-I B-R-V-Y-N-I B-R-V-Y-U-I 8/2 88/ Alexa Fluor 79 Alexa Fluor 7 CF79 PF8

19 B-R-V CytoFLEX CytoFLEX LX CytoFLEX S CytoFLEX Blue-Red- Violet- Yellow Green-Near UV-Infrared B-R-V-Y- N-I Blue-Red- Violet- Yellow Green-UV- Infrared B-R-V-Y- U-I Blue-Red- Violet- Yellow Green B-R-V-Y Blue-Red- Violet-Near UV B-R-V-N Blue-Red- Violet- Infrared B-R-V-I Blue-Violet- Yellow Green-Near UV B-V-Y-N Blue-Red- Violet 7 nm mw nm mw 8 nm mw - nm 8 mw 1 nm mw nm 2 mw nm mw Si- Avalanche Photo Diode detector arrays FAPD Fiber Array Photo Detector) QC FCS Area Height Width 1 7 FITC MESF PE MESF 88nm rcv 88 nm. m nm.2 m, / Slow L/min Medium L/min Fast L/min 2 L/min ml 12 7 mm 1. ml 2 ml * 9 U V Semi Automatic L 2 L V U L 1.% *.% Deep Clean CytExpert / / API (Application Programming Interface)* QC Pass/Fail L-J FCS2. FCS. 7 W x D 1 H mm 79 kg 2 W x 2 D H mm 2. kg 1 2W 1 27 Kaluza * 19

20 Join the Resolution REVOLUTION Beckman Coulter Beckman Coulter Beckman Coulter, Inc. CE L

Amnis TM CellStream TM CellStream TM

Amnis TM CellStream TM CellStream TM Amnis TM CellStream TM Amnis TM CCD FITC PE 5 96 7 CFR Part FDA TM CellStream Luminex Time Delay Integration TDI CellStream. 7... CCD CellStream Amnis CCD CellStream