Efficient, flexible, transformative flow cytometer with up to 14 colors

The Attune NxT Flow Cytometer is an advanced cell analyzer in which acoustic focusing fluidics is the key to both high sensitivity and high throughput.


Features

The Attune NxT Flow Cytometer shares common features with the Attune CytPix model.

Acoustic focusing for sensitivity

Fast, accurate acquisition

Rare event detection

Designed for flexibility

Novel optical design

Clog-resistant technology

Powerful, intuitive software

Biosafety hood compatible

For our imaging-enhanced flow cytometer, check out the Attune CytPix model.


Flow cytometry applications

Almost any standard flow cytometry application can benefit from the high sensitivity and throughput of the Attune NxT Flow Cytometer. See our Sample Data page for examples spanning infectious disease research, fluorescent proteins, CRISPR gene editing, immuno-oncology research, microbiology, plant ploidy, platelets, stem cells, and T cells.

 

One classic flow cytometry application is immunophenotyping. Flow cytometry is the method of choice for identifying cells within complex heterogeneous populations, as it allows for multiparameter analysis of thousands to millions of cells in a short time. Strong signal separation in the Attune NxT Flow Cytometer shows excellent separation of cell populations into subsets for immunophenotyping. A wide range of reagent choices—as well as the system’s automated compensation module, four spatially separated lasers, and 14 color choices—help to simplify multicolor panel design.

Thirteen color immunophenotyping using Attune NxT

Gating strategy for 13-color immunophenotyping analysis of stained human whole blood using a stain/lyse protocol. Human whole blood cells were stained as described in the application note and acquired and analyzed on the Attune NxT Flow Cytometer. (A) Dead cells were excluded from the analysis by gating on live (propidium iodide) cells in a dot plot. (B) CD45+ gating was used to select the leukocyte population from the lysed whole blood. (C) Lymphocytes and monocytes were identified based on forward and side scatter profiles. (D) Monocytes are found above lymphocytes on the scatter plot and express both CD14 and CD33. (F) Within the lymphocyte gate, immune cells can be subdivided based on their expression of CD3 (T cells), CD19 (B cells), or neither (NK cells). (E) B cells can be further characterized by HLA-DR and CD45RA expression. (G) T cells can be further subdivided into CD4+ (T helper cells) and CD8+ (cytotoxic T cells) subpopulations, while (J) regulatory T cells express CD4 and CD25. (H, K) CD62L identifies naive (TN) CD4 and CD8 T cells, while HLA-DR is expressed by activated T cells (TA). (I) Finally, NK cells lack B cell and T cell markers (CD19CD3) and express CD56.


Videos and Demos


Laser and detector configurations

Lasers Laser configuration Lasers included, No. of detection channels Total detection channels* Category Number
Violet, 405nm Blue, 488nm Yellow, 561nm Green, 532nm Red, 637nm
1 Blue
4 6 A24864
2 Blue/green
3 4 9 A28995
Blue/yellow
3 4 9 A24861
Blue/red
4 3 9 A24863
Blue/violet
4 4 10 A24862
Blue/violet 6
6 3 11 A29002
3 Blue/green/red
3 4 3 12 A28997
Blue/red/yellow
3 4 3 12 A28993
Blue/green/violet
4 3 4 13 A28999
Blue/violet/yellow
4 3 4 13 A24859
Blue/red/violet
4 4 3 13 A24860
Blue/red/violet 6
6 3 3 14 A29003
4 Blue/red/violet/green
4 3 4 3 16 A29001
Blue/red/yellow/violet
4 3 4 3 16 A24858
Blue/red/yellow/violet 6
6 2 3 3 16 A29004

Specifications

Optics: Fluorescence detection

Laser power

 

 

Laser

Wavelength (nm)

Beam-shaping optics (BSO)* (mW)

Diode power** (mW)

Violet

405

50

100

Blue

488

50

100

Green

532

100

140

Yellow

561

50

100

Red

637

100

140

* 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); user-changeable, 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 turns on when acquiring samples; reports hours of usage

Flat-top laser specified at the flow cell

Coefficient of variation (CV) <3% over the width of the flat-top laser

Upgradable

Convenient field changes

Fluidics

Flow cell

Quartz cuvette gel coupled to 1.2 numerical aperture (NA) collection lens, 200 x 200 μm

Sample analysis volume

20 μL–4 mL

Custom sample flow rates

12.5–1,000 μL/min

Sample delivery

Positive-displacement syringe pump for volumetric analysis

Sample tubes

Accommodates tubes from 17 x 100 mm to 8.5 x 45 mm

Fluid-level sensing

Active

Standard fluid reservoirs

1.8 L focusing fluid tank, 1.8 L waste tank, 175 mL shutdown solution tank, and 175 mL wash solution tank

Fluid storage

All fluids stored within instrument

Extended fluidics option

Configuration for 10 L fluid

Nominal fluid consumption

1.8 L/day

Automated maintenance cycles

≤15 min start-up and shutdown—deep clean, sanitize, and debubble modes

Performance: Fluorescence detection

Fluorescence sensitivity

≤80 molecules of equivalent soluble fluorochrome (MESF) for FITC, ≤30 MESF for PE, ≤70 MESF for APC

Fluorescence resolution

CV <3% for the singlet peak of propidium iodide–stained chicken erythrocyte nuclei (CEN)

Data acquisition rate

Up to 35,000 events/sec, 34 parameters, based on a 10% coincidence rate per Poisson 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 in lysed 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—0.1 μm on side scatter under following conditions: Using an Attune NxT Flow Cytometer with standard 0.5 mm blocking configuration, an Invitrogen Attune NxT 488/10 Filter (Cat. No. 100083194), and Attune Focusing Fluid (Cat. No. 4488621, 4449791, or A24904) that has been passed through a 0.025 µm filter

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

Hierarchical gating with the ability to derive gates

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 map settings, and compensation settings optimized and defined previously

Tube-to-plate conversion

One-click transition from tubes to plates and vice versa; no disassembly, no additional 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 copy and 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 count

Quality and regulatory

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 performance to specifications

Quality management system

Manufacturing standards comply with the requirements of ISO 13485:2003

Robust installation specifications

Units installed by engineer; preplanning checklist, delivery, and installation; and performance validation compliance with standardized procedure

Regulatory status

For Research Use Only

Computer

Software requirements

Invitrogen Attune Cytometric Software

Monitor

23 in. flat panel (1,920 x 1,080 resolution); dual-monitor capability

Computer

Minitower desktop

Operating system

Microsoft™ Windows™ 10 64-bit

FCS format

FCS 3.1, 3.0

Processor

Intel™ Core™ i7 processor

RAM

32 GB

Hard drives

2 x 2 TB SATA 3.0 Gb/s, 8 MB data burst cache; controller RAID1, integrated

Installation requirements

Electrical requirements

100–240 VAC, 50/60 Hz, <150 W
Thermo Fisher Scientific certifies that the Attune Flow Cytometers conform to relevant directives to bear the CE mark. The instrument also conforms to the UL and CAN/CSA general requirements (61010.1). The Attune Flow Cytometers are Class I laser products 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)

For Research Use Only. Not for use in diagnostic procedures.