SPECIFICATION SHEETAttune NxT Flow CytometerAttune NxT Flow CytometerOn the quest for significant breakthroughs, neversettle for averageRun samples faster and achieve greater resolution withlittle fear of sample loss due to clogging. The Invitrogen Attune NxT Flow Cytometer combines precision withperformance in a true benchtop flow cytometer with up to4 lasers and 16 parameters of detection. The Attune NxTFlow Cytometer was developed with the goal of removingbarriers associated with flow cytometry and enablingnew research. This includes bringing the power of flowcytometry to the analysis of more sample types, includingcancer cells. The addition of the Invitrogen Attune NxTAutosampler enables significantly faster high-throughputscreening on a multiparametric platform.The system offers: Time savings—up to 10x faster data acquisition speeds Reduced clogging—engineered for even large or stickycell types Efficient protocols—rapid no-wash, no-lyse sampleprep helps reduce protocol time and minimize cell loss Robust software—developed to offer user-focusedfunctionality with many automated, user-definable, andadministrative features Reduced need for compensation—spatiallyseparated lasers Elegant fluidics design—readily accessible, easy to fill,and require little-to-no maintenance Rare-event detection—technology for identifyingvery rare subpopulations of cells with high efficiencyand certainty Flexibility—choice of lasers and parameters Multiple configurations—for a broad arrayof applications

Instrument specificationsOptics Laser power (as shown in table below)LaserWavelength (nm)Beam-shaping optics (BSO)* (mW)Diode power** 56150100Red637100140* Amount of measured usable laser power after light has gone through the beam optics and shaping filters.** Vendor-specified theoretical maximum. Laser excitation: Optimized excitation for minimized stray laser-line noise and losses to reflection Laser profile: 10 x 50 μm flat-top laser providing robust alignment Emission filters: Up to 14 color channels with wavelength-tuned photomultiplier tubes (PMTs); userchangeable, keyed filters Laser separation: 150 μm Optical alignment: Fixed alignment with prealigned welded fiber; no user maintenance required Onboard thermoelectric cooler: No warm-up delay; fiber isn’t affected by on/off Simmer mode: Instant on/off reduces usage and/or aging by 10x; only keep it “on” when acquiring samples;reports hours of usage Flat top specified at the flow cell: Coefficient of variation (CV) 3% over width of flat top Upgradable: Convenient field changesFluidics PerformanceFlow cell: Quartz cuvette gel coupled to 1.2 numerical aperture (NA) collection lens, 200 x 200 μmSample analysis volume: 20 μL to 4 mLCustom sample flow rates: 12.5–1,000 μL/minSample delivery: Positive-displacement syringe pump for volumetric analysisSample tubes: Accommodates tubes from 17 x 100 mm to 8.5 x 45 mmFluid-level sensing: ActiveStandard fluid reservoirs: 1.8 L focusing fluid tank, 1.8 L waste tank, 175 mL shutdown solution tank, and175 mL wash solution tankFluid storage: All fluids stored within instrumentExtended fluidics option: Configuration for 10 L fluidNominal fluid consumption: 1.8 L/dayAutomated maintenance cycles: 15 min startup and shutdown—deep clean, sanitize, and debubble modes Fluorescence sensitivity: 80 molecules of equivalent soluble fluorochrome (MESF) for FITC, 30 MESF forPE, 70 MESF for APC Fluorescence resolution: CV 3% for the singlet peak of propidium iodide–stained chicken erythrocytenuclei (CEN) Data acquisition rate: Up to 35,000 events/sec, 34 parameters, based on a 10% coincidence rate perPoisson statistics Maximum electronic speed: 65,000 events/sec with all parameters Carryover: Single-tube format: 1% Forward and side scatter sensitivity: Able to discriminate platelets from noise Forward and side scatter resolution: Optimized to resolve lymphocytes, monocytes, and granulocytes inlysed whole blood Forward scatter: Photodiode detector with 488/10 nm bandpass filter Side scatter: PMT with default 488/10 nm bandpass filter; optional 405/10 nm bandpass filter Fluorescence detectors: 14 individual detectors Electronic pulse: Measured area, height and width pulse for all detectors Violet side scatter resolution: Can be configured for violet side scatter to better resolve particles from noise Minimum particle size: 0.2 μm on side scatter using submicron bead calibration kit from Bangs Laboratories

Instrument specifications (continued)Software Compensation: Full matrix—automated and manual modes, on-plot compensation tools for fine adjustment;use of tubes and wells Flow rate: Precise flow rate control via software; no hardware adjustments Live streaming: Live update of statistics during acquisition of events up to 35,000 events/sec Overlays: Comparative analysis between samples; 3D view Sample recovery: System able to return unused samples Concentration: Direct concentration measurement without use of counting beads Software layout: Fully customizable for each user account Bubble detection technology: Stops automated run to preserve sample integrity Maximum single-event file: 20 million with option to append Heat map: Set up for definition of plate layout; screening view for analysis for tubes and plates Threshold: Up to 4 individual thresholds with user option to apply Boolean logic Gating: Hierarchal gating with the ability to derive gates Smartgate labeling: Option to annotate quad gate names based on fluorophore and target names Voltage: User adjustable Window extensions: User adjustable Area scaling factor (ASF): User adjustable Acquisition settings: Documented in FCS files and maintained upon import Templates: Create from existing experiments—instrument settings, workspaces, run protocols, heat mapsettings, and compensation settings optimized and defined previously Tube-to-plate conversion: One-click transition from tubes to plates and vice versa; no disassembly, noadditional QC, no reboot required for conversion between plates and tubes Graphics resolution: Publication-quality images; support for TIF, PNG, BMP, JPG, GIF, and EMF; quickly copyand paste plots to any external application (e.g., Microsoft PowerPoint software) User account administration: Administrative creation of individual user accounts with designated roles,advanced setting permissions, management of individual accounts, user time tracking, and sample countQuality andregulatory Instrument tracking: Automated daily baseline and performance test with Levey-Jennings plots Warranty: 1 year Production verification testing: Each instrument is tested and verified for assembly integrity and performanceto specifications Quality management system: Manufacturing standards comply with the requirements of ISO 13485:2003 Robust installation specifications: Units installed by engineer; preplanning checklist, delivery, andinstallation; and performance validation compliance with standardized procedure For Research Use OnlyDatamanagement Software requirements: Invitrogen Attune NxT SoftwareMonitor: 23-inch flat panel (1,920 x 1,200 resolution); dual-monitor capabilityComputer: Minitower desktopOperating system: Windows 7 64-bitFCS format: FCS 3.1, 3.0Processor: Intel Core i7 processorRAM: 16 GBHard drives: 80 GB or larger and 250 GB redundant array of independent disks (RAID)–compatible hard drives

Instrument specifications (continued)Installationrequirements Electrical requirements: 100–240 VAC, 50/60 Hz, 150 WThermo Fisher Scientific certifies that the Attune NxT Flow Cytometer conforms to relevant directives to bearthe CE mark. The instrument also conforms to the UL and CAN/CSA general requirements (61010.1). TheAttune NxT Flow Cytometer is a Class I laser product per Center for Devices and Radiological Health (CDRH)regulations and EN/IEC 60825. Heat dissipation: 150 W Temperature operating ranges: 15–30 C (59–86 F) Operating humidity: 10–90%, noncondensing Audible noise: 65 dBA at 1.0 m Instrument size (H x W x D): 40 x 58 x 43 cm (16 x 23 x 17 in.), including fluid bottles Weight: 29 kg (64 lb) Available configurations (as shown in table below)LasersLaserconfiguration(Cat. No.)Violet405 nmBlue488 nmYellow561 nmGreen532 nmRed637 nmTotaldetectionchannels*1Blue(A24864)Available asupgrade4Available asupgradeAvailable asupgradeAvailable asupgrade62Blue/green(A28995)Available asupgrade3–4Available asupgrade9Blue/yellow(A24861)Available asupgrade34–Available asupgrade9Blue/violet(A24862)44Available asupgradeAvailable asupgradeAvailable asupgrade10Blue/red(A24863)Available asupgrade4Available asupgradeAvailable asupgrade39Blue/green/red(A28997)Available asupgrade3–431243–4Available asupgrade13Available vailable asupgrade13Blue/red/violet(A24860)44Available asupgradeAvailable asupgrade313Blue/red/violet/green (A29001)43–4316Blue/red/yellow/violet d/yellow(A28993)4* Includes forward scatter (FSC) and side scatter (SSC).

Attune NxT AutosamplerTo improve experimental workflow, the Attune NxTAutosampler is a high-throughput sampler option for theAttune NxT Flow Cytometer. The system offers: Broad compatibility—compatible with many differentplate formats, including 96-well, 384-well, anddeep-well plates Intelligent probe design—minimizes clogging Auto cleaning—performs automated cleaning when theinstrument is shutting down Consistent data—designed to provide minimal variationregardless of sampling method (tube vs. plate) andcollection rate Mixing by aspiration—mixing sample by aspirationinstead of shaking helps ensure homogeneity of thesample and maintains cell viability One-click transition—plate and tube compatibility: nodisassembly, no additional QC, no reboot required forconversion between plates and tubesInstrument specificationsPerformance High-throughput mode acquisition time: 42 min for 96-well plate, 180 min for 384-well plate (using onerinse and one mix, and full analysis of a 40 μL sample) Carryover: 0.5% in plate loader format (standard mode, 2 wash cycles); multiple-rinse capability forultralow carryover Sample mixing: Mixing optimized to preserve cell viability; mixing cycles optimized to sample analysis volume Mixing method: Each well mixed via full aspiration (no shaking) Wash cycle: User-defined number of wash cycles, dependent on plate-processing protocol and time toacquire plates Minimum dead volume (single draw): 30 μL for 12.5–200 μL/min, 50 μL for 50–1,000 μL/min Sample window: Protectively coated window allows visibility to well progress while preventing exposure toambient light during acquisition Auto-calibration: Regular, 30-day interval, system-initiated functionFluidics Plate and tube compatibility: One-click transition from tubes to plates and vice versa; no disassembly, noadditional QC, no reboot required for conversion between plates and tubes Compatible plate types: 96 deep well (flat, round, and V-bottom), 96-well standard depth (flat, round, andV-bottom), 384-well standard depth (flat, round, and V-bottom), 384 deep well (flat, round, and V-bottom) Cleaning cycles: Automated daily and monthly cleaning protocols Fluidics requirements: 800 mL total of onboard fluid tanks, capable of running four 96-well plates Extended fluidics option: Optional external fluid tank with 10 L fluid capacity

Instrument specifications (continued)Installationrequirements Size (W x D x H): 40 x 29 x 29 cm (16 x 11 x 11 in.) Space requirements:–– Minimum width: 40 cm (15.8 in.); when attached to the Attune NxT Flow Cytometer, the total width is 167 cm(65.8 in.)–– Minimum depth: 58.5 cm (23.1 in.) provides 43.2 cm (17.1 in.) for the cytometer unit, a 10.2 cm (4 in.) ledge infront of the unit to place fluidics bottles, and 6.5 cm (2.5 in.) behind the unit for ventilation–– Minimum clear height: 74 cm (29 in.) above the mounting Mounting: Side Weight: 16 kg (35 lb) Operating range (environmental conditions): 15–30 C (50–95 F) Operating humidity: 80% noncondensing Electrical requirements: 100–240 VAC, 50/60 Hz, 300 WComparing flow cytometers among manufacturersThe purpose of this specification sheet is to help youidentify design attributes of the Attune NxT Flow Cytometer,such as performance, size, environment, and software, todetermine if the instrument is a good fit for your research.The specifications can be used as a basis for comparison,helping you assess the value of different instruments forthe price. This specification sheet is also a guide to theexpected performance. For this reason, you should havea good understanding of the stated values and how theypertain to your intended use of the instrument. When usingthis document as a comparison guide across platforms,be aware that there are many performance values thatappear comparable across instruments but in reality arequite different. A specification is derived from a specific testor calculation, but these tests are not standardized acrossinstrument developers and may be misleading in a side-byside comparison.Education, service, and supportFlow cytometry learning center: Find a rangeof targeted resources for flow cytometry, fromwebinars that cover the basics of this technology toapplication notes that demonstrate the utility of flowcytometry for high-content analysis applications. Go Hands-on, in-lab training is available. Customertraining centers are also available in selected countries.Service and support: Reliable global technical support,experienced field service engineers, and flow cytometryspecialists are available for assistance.Remote monitoring and diagnostics service: TheSmart Monitor tool is a real-time, remote instrumentperformance monitoring service* that incorporates aninstrument management dashboard, remote diagnostics,and aggregated performance data tracking.Service packages at a glanceAB Complete PlanAB Assurance PlanAB Maintenance Plus PlanOn-site response timeGuaranteednext business day**Guaranteed2 business days**Target2 business days**Scheduled planned maintenance Application technical support Instrument operatingsoftware updates On-site service—labor, parts, travel 10% discountPhone and email access to engineersin the remote service center Computer coverage Remote instrumentmonitoring diagnostics On-site application consulting Qualification service * Subject to Internet connectivity.** Availability limited in some geographic areas.

Workflow portfolio to support your research goalsAt Thermo Fisher Scientific, we are committed toaccelerating your research by providing a comprehensivesuite of solutions for the analysis of cells and theirfunctions. Behind this commitment is a dedicated andtalented team of scientists developing and supportingour innovative instrumentation and products such as theAttune NxT Flow Cytometer, Invitrogen eBioscience flowcytometry antibodies, and Invitrogen cell health reagents.Antibodies—The Invitrogen portfolio offers over15,000 flow-specific conjugated antibodies with multiplefluorophore options, including the new Invitrogen SuperBright violet-excitable polymer dyes. Our antibodies enableyou to build and expand your panels as your researchdemands.Buffers—The use of appropriate buffers is crucial to thesuccess of your flow cytometry experiments. We offer awide variety of buffers to suit your research needs whetheryour experiment calls for extracellular, intracellular, and/ornuclear staining.Flow support products—Compensation beads areessential when performing flow cytometry using multiplechannels, markers that are poorly expressed, or whensample is limited. The one-vial, one-drop approachenabled by our Invitrogen OneComp and UltraCompeBeads products provides remarkable ease of use forcompensating your antibody.We are focused on advancing meaningful discoveriesand partnering to make tools for cellular analysis widelyaccessible, affordable, and powerful for all life scientists.Reagents—At the forefront of invention and developmentof fluorescent probes for over 40 years, we offer acomprehensive variety of functional assays for studying cellviability, apoptosis, cell cycle, and cell proliferation.Find your reagents at out more about our flow cytometry products andservices at Research Use Only. Not for use in diagnostic procedures. 2017 Thermo Fisher Scientific Inc. All rights reserved. Alltrademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Microsoft, PowerPoint, and Windowsare trademarks of Microsoft Corp. Intel Core is a trademark of Intel Corp. Bangs Laboratories, Inc. is a division of Polysciences, Inc.COL31763 0517

Attune NxT Flow Cytometer combines precision with performance in a true benchtop flow cytometer with up to 4 lasers and 16 parameters of detection. The Attune NxT Flow Cytometer was developed with the goal of removing barriers associated with flow cytometry and enabling new research. This