capabilities today. Flexibility for tomorrow.

Transcription

1 capabilities today. Flexibility for tomorrow. NEW CellStream benchtop flow cytometry system with Amnis detection technology inside The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.

2 Capabilities Today. Flexibility for Tomorrow. Unparalleled combination of FLEXIBILITY & PERFORMANCE The CellStream flow cytometer is a new benchtop system that offers unparalleled capability, sensitivity, and expandability at an accessible price. Contains patent-protected camera technology unique to our state-of-the-art Amnis flow cytometers Single tube and 96-well plate sampling CFR Part -enabling features Automated daily system calibration Unique Event Gallery for visual sample verification High sensitivity Highly flexible Patented optics system Highthroughput acquisition Intuitive software Enabled by a single CCD detector that replaces PMTs MESF < FITC; MESF <5 PE Excellent small particle detection Resolves complex cell populations Fully field upgradeable to 7 lasers provide up to detection channels

3 Inside the CellStream System Our patented time delay integration (TDI) and camera technology delivers sensitivity and expandability beyond what is possible with traditional flow cytometers. CellStream system architecture. Up to 7 lasers are focused in discrete locations.. Hydrodynamically focused cells pass through the laser-illuminated region. Fluorochromes bound to the cells are excited and emit into the collection system. Fluorescence is collected and directed toward an intermediate image plane.. The filter stack decomposes each of the four discrete vertical positions in the intermediate image plane into separate channels of data.. All channels fit efficiently onto a CCD (charge-coupled device) array. CellStream system s sensor contains multiple discrete collection fields using the same CCD as patented Amnis technology. EMDMillipore.com/cellstream

4 Capabilities Today. Flexibility for Tomorrow. STREAM THE POWER OF SENSITIVITY High sensitivity fluorescence detection Fluorescence sensitivity of the CellStream flow cytometry platform was evaluated using industry standard 8-peak Spherotech rainbow calibration beads. The data demonstrate high fluorescence sensitivity of the CellStream system: All 8 peaks are clearly resolved on every detection channel Low MESF (Molecules of Equivalent Soluble Fluorochrome) values determined: MESF < FITC; MESF <5 PE CellStream Instrument Channels 56/5 58/6 58/ 6/ 7/87 77/56 75 nm e e e5 e6 56/ e e e5 e6 58/6.5.5 e e e5 58/ CellStream Lasers 5 nm 88 nm e e e5 e6 56/5 e e e5 e6 58/ e e e5 e6 58/6 e e e5 e6 58/ e e e5 e6 58/ e e e5 e6 6/ e e e5 e6 6/ e e e5 e6 7/87 e e e5 e6 7/ e e e5 77/ nm e e e5 e6 58/ e e e5 e6 6/ e e e5 e6 7/87 e e e5 e6 77/56 6 nm e e e5 e6 7/ e e e5 77/56 High sensitivity submicron particle detection The CellStream flow cytometer clearly detects and discriminates particles as small as. μm. The figure shows the acquisition of Megamix-Plus FSC size beads containing, 5, and 9 nm fluorescent beads in a known ratio of ::. Instrument settings: 7 mw SSC, % FSC, and mw 88 nm; slow speed. Side Scatter (SSC) e+5 e+ e+ e+ e+ Forward Scatter (FSC)

5 STREAM SUPERIOR DETECTION OF SMALL PARTICLES Only recently has the importance of extracellular vesicles (EVs) as key mediators of intercellular communication been appreciated. EVs are membrane-derived structures that include exosomes, microvesicles and apoptotic bodies. In this study, RBC-derived EVs were stained with anti-cd5ab-pe and/or anti-cd5ab-apc. Control samples were collected for antibody only, PBS, and RBC EVs labeled with anti-cd5ab-pe and anti-cd5ab-apc incubated with Triton-X (TX). (A) Initial gate to identify potential EVs. Using this gate, (B) PE+ and (C) APC+ events were identified. (D) Chart showing PE+ and APC+ objects/ml for the control samples and the CD5ab-PE and anti-cd5ab-apc labeled samples. A Population: All B Population: Potential EVs 5 8 SSC 5 5 Frequency 6 PE+99.% Potential EVs 87.% 5 FSC 5 5 e+ e+ PE-Intensity C Population: Potential EVs D 5 Frequency APC-Intensity APC+98.7% e+ e+ Objects/mL Antibody EV + PE EV+PE/APC EV+APC Sample Condition PBS EV+PE/APC+TX EMDMillipore.com/cellstream 5

6 Capabilities Today. Flexibility for Tomorrow. STREAM THE POWER OF VERSATILITY CellStream flow cytometer enables cell researchers to obtain reproducible, multi-parametric single cell data for a wide variety of applications. Immunological Phenotyping -Color Assay In this example, a -laser CellStream system accurately resolves different fluorochromes within a single assay. Below, different immune cell populations were clearly resolved from one another within a sample of PBMCs. Side Scatter Side Scatter Side Scatter CD BV CD5 BV65 CD5 APC-Cy7 CD5+ Side Scatter Side Scatter Forward Scatter CD PE-Cy5.5 CD- CD+ CD+ CD8+ CD5RA APC CD5RA APC CD+ or CD8+ CD9 BUV96 CD8 BV5 CD8 CD CD6L FITC CD6L FITC CD6 + CD56 PE CD PE-Cy7 Staining protocol 5 μl sample* of PBMCs were stained for minutes with the following fluorochromes (5 μl each): CD9 BUV 96 (B Cells) CD6 + CD56 PE (NK cells) CD BV (Monocytes) CD PE-Cy5.5 (T Cells) CD8 BV 5 (CD8 T Cells) CD PE-Cy7 (CD T Cells) CD5 BV 65 (Granulocytes) CD5RA APC (Naïve and Memory T cells) CD6L FITC (Naïve and Memory T cells) CD5 APC-Cy7 (Lymphocytes) After staining, samples were washed once, resuspended in Guava Antibody Dilution Buffer, and acquired on the CellStream system. A total of, lymphocyte events were acquired. *Approx million white blood cells, depending on donor. 6

7 A Fully Configurable System CellStream systems are made to order. Build an instrument specific for your needs from the available lasers below. All systems come standard with: AutoSampler for 96-well plates Single tube sampler 88 nm laser Excitation & Emission Capabilities of the 7-Laser CellStream Flow Cytometer CellStream Lasers CellStream Instrument Channels 56/5 58/6 58/ 6/ 7/87 77/56 75 nm AF5 BUV95 BUV96 BUV56 QDot65 BUV66 BUV85 5 nm BV Cascade Blue BV5 QDot565 BV65 BV65 BV786 FITC AF88 PE PE-TR PE-Cy5.5 PE-Cy7 5 nm PE mcherry PE-TR PE-Cy5.5 PE-Cy7 56 nm PE mcherry PE-TR PE-Cy5.5 PE-Cy7 APC Cy5 APC-Cy7 APC-AF75 88 nm 6 nm 7 nm Cy7 CF75 Inside the 7-Laser CellStream System EMDMillipore.com/cellstream 7

8 Capabilities Today. Flexibility for Tomorrow. STREAM THE CONFIDENCE OF INTUITIVE SOFTWARE Integrated software provides an intuitive and easy-to-use interface, enabling you to focus on your experiments and your data. Software includes CFR Part -enabling features for quality control and data integrity, essential in regulated environments. Load & Record Tubes or plates Simple & customizable AutoSampler set up Toolbar Quickly define experiments, view Event Gallery or access other frequently-used parameters Startup/Shutdown/System Status One click: Initialization & daily cleaning with on-board fluidics Calibration & testing (laser alignment, dark current, flow core position, flow core stability, channel alignment, laser power) Sample Listing Settings Record by count, volume or time Intuitive control of instrument, experiment and plotting parameters, and thresholds Pop-up fluorochrome chart for easy channel identification Display & Analysis Full suite of data display and analysis tools (histograms, dot plots, density plots, overlays, dot plot backgating, multi-file analysis, ) Streamlined acquisition of compensation files Export statistics or create customized PDF reports 8

9 A unique Event Gallery feature of CellStream Acquisition Software allows for population verification, aids in troubleshooting and resolves doublets. Real-time Event Gallery Low resolution images of your cells in flow Provides verification of suspected populations Aids in troubleshooting Unlike any other non-imaging flow cytometer Singlets Doublet Discrimination Aspect ratio feature allows for visual confirmation Clear resolution between singlet, doublet and aggregate events Calculated for each channel Aspect Ratio e+ e+ 6e+ 8e+ e+5 FSC Doublets Aggregates EMDMillipore.com/cellstream 9

10 Capabilities Today. Flexibility for Tomorrow. Instrument Service Plans To help you get the most out of your CellStream flow cytometry system, our worldwide service organization offers a variety of service plans to support your individual needs and maintain the longevity of your instrument. Our service agreements are structured yet flexible, so you can select the level of hardware, application and software support you prefer. Advantages of maintaining a service plan: Best-in-class service support maintains optimal performance, enabling high-quality data Planned instrument maintenance reduces overall service costs Service plans are the best protection for your instrument investment and its long-term operation Instrument Service Plan Options Service Plan Features* One-Year Warranty Service Total Plan Travel & Labor Expenses Coverage of Parts Service Advanced Plan Service Essential Plan Preventative Maintenance Visit Factory Trained & Authorized Service Engineers Technical Support Services Priority Scheduling One visit for instrument diagnostics or basic repairs *No service contract is required for one-time service requests. Our highly-qualified field application and instrument specialists also provide: Support by or phone On-site instrument training On-site scientific applications support For more information on our comprehensive range of service and support agreements, please contact your sales representative or visit EMDMillipore.com/cellstream.

11 System Performance Parameter Fluorescence sensitivity CV* (precision) <% Number of channels Number of lasers -7 Available lasers Camera-enabled morphology parameters Event rate Flow rates Scatter resolution Dynamic range System size (W D H) Field upgradeable Sample formats Absolute Cell ing g Performance MESF < FITC MESF <5 PE Up to ( fluorescent, plus FSC, SSC) 75, 5, 88, 5, 56, 6, 7 nm (area; aspect ratio; raw max. pixel), cells/second.66 µl/min (Low speed/high sensitivity).6 µl/min (High speed) FSC < nm from 5 nm SSC < nm from 785 nm 7 decades mm Yes Single tube or 96-well plate Yes *Coefficient of Variation using Chicken Erythrocyte Nuclei (CEN) Ordering Information Description CellStream Base System with 88 nm laser and AutoSampler CellStream Four-Laser System with 88 nm, 6 nm, 5 nm, 56 nm lasers and AutoSampler CellStream Option 75 nm Laser CellStream Option 5 nm Laser CellStream Option 5 nm Laser CellStream Option 56 nm Laser CellStream Option 6 nm Laser CellStream Option 7 nm Laser CellStream Software Multi Access CellStream Calibration Reagent CellStream On-Site Training CellStream Installation CellStream IQOQ Document Cat. No. CS-96 CS-96 CS-75 CS-5 CS-5 CS-56 CS-6 CS-7 CS- CS- CS-5 CS-6 CS-65 Absolute Cell ing Yes EMDMillipore.com/cellstream

12 MilliporeSigma Summit Drive Burlington, MA 8 To place an order or receive technical assistance In the U.S. and Canada, call toll-free For other countries across Europe and the world, please visit: EMDMillipore.com/offices For Technical Service, please visit: EMDMillipore.com/techservice EMDMillipore.com/cellstream 8 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. MilliporeSigma, the Vibrant M, CellStream, Guava, Amnis, Service Total, Service Advanced and Service Essential are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources. Lit. No. MS_BR8EN Ver.. 8-6/8