Features common to all Attune flow cytometers

The Invitrogen Attune flow cytometer family offers a range of advanced features including high-speed sample acquisition, clog-resistant fluidics, flexible optical design, and exceptional sensitivity, setting a new standard in flow cytometry and empowering researchers to make their discoveries faster and with greater precision.

Try our instrument configurator tool to design an Attune NxT or CytPix system to fit your research needs.


Acoustic focusing for sensitive analysis

Attune Flow Cytometers combine ultrasonic waves like those used in medical imaging with hydrodynamic forces to precisely position cells into a single, focused line in the central axis. Enabling cells to be tightly focused at the point of laser interrogation allows the system to collect more photons, helping to ensure data quality regardless of the sample-to-sheath ratio.

Acoustic focusing

Acoustic focusing (left) precisely positions cells in tight alignment even at higher sample rates resulting in less signal variation and better data quality. Traditional hydrodynamic focusing (right) widens the sample stream core at high sample rates that results in signal variation and compromised data quality.

Effect of acoustic focusing on cells in sample stream.

Fast sample acquisition speed

Acoustic focusing also lets your lab rapidly acquire high-quality data. Users can achieve sample throughput rates of 12.5 µL/min or 1,000 µL/min, up to 10 times faster than traditional hydrodynamic focusing systems and acquisition speeds of 35,000 events per second. This means processing all samples—including low-concentration and precious samples—more quickly and accurately with minimal loss in quality.

Time to acquire 1 million events

Rapid data acquisition

 

Compares the time required to acquire 1,000,000 events over three competitor instruments running at maximum sample rates.

Acoustic focusing minimizes variation regardless of the sample rate, so you don't have to make the tradeoff between throughput and sensitivity. This is demonstrated in the cell cycle analysis example below, where it's critical to precisely detect differences in fluorescence intensity between multiple cell populations.

Consistent data collection independent of sample flow rates between 12.5 and 1000 microliters per minute Consistent data collection independent of sample flow rates between 12.5 and 1000 microliters per minute

Minimal data variation

 

Consistent results were achieved at high sample rates using Jurkat cells fixed and stained with propidium iodide, treated with RNase and analyzed at a concentration of 1 x 106 cells/mL. The coefficient of variation (CV) of cells in the G0/G1 and G2/M phases remain consistent, even at the highest sample rate of 1,000 μL/min.


Clog resistant fluidics

To prevent clogging and allow for volumetric analysis, the Attune Flow Cytometers use a positive displacement syringe pump to control sample volume. The system can perform volumetric cell counts in a known volume (gated or total events) and can easily gate out dead cells to count live cells only in a live/dead analysis. Unlike traditional cytometers that operate at a maximum pressure of 15 PSI, Attune Flow Cytometers control pressure at 75 PSI, reducing the likelihood of cell-cell interactions and resulting in clog resistance.

 

Syringe pump aspirating sample from sample tube.

Volumetric sample entry

 

Using positive displacement, the syringe pump works like a pipette. Tension is created with the plunger pushing down (A). The tension is released as the plunger rises with the sample (B). Clogging is less likely to occur under this high and controlled pressure environment.


Flexible optical design

Attune Flow Cytometers are configurable with up to 4 spatially separated lasers and 16 parameters. Spatial separation provides flexibility for multicolor panel design and streamlines compensation.  The system offers superior speed with acquisition rates of up to 35,000 events per/second with high sensitivity to meet a range of research requirements.

 

High sensitivity distinguishes between dim signals and background, resulting in less variation and better signal separation. Fluorescent resolution coefficient variation is less than 3% for a single peak, and predicted MESF is ≤80 (FITC), ≤30 (PE), ≤70 (APC). Sensitivity in comparison to competitive systems is described below.

Sensitivity measurements across flow rates

Fluorescent microspheres (Spherotech Rainbow 3.2 μm) were run on a high-end conventional flow cytometer (A) and on the Attune NxT Flow Cytometer (B and C) using a 561 nm laser and 610/20 (A) or 610/15 (B and C) emission filters. The conventional cytometer was run using the highest sensitivity setting (~12.5 μL/min). The Attune NxT Flow Cytometer was run at 12.5 μL/min (B), which is equivalent to the traditional flow cytometer and 500 μL/min (C; 40x more sample). The Attune NxT Flow Cytometer results were equal to or better than those from the conventional flow cytometer, even at the highest flow rate.

The compact size of the Attune Flow Cytometer also provides the flexibility of using it within a biosafety hood. This helps avoid contamination or infection when working with hazardous or unknown samples.


Lasers designed for stability

Attune Flow Cytometers feature a novel optical design that delivers first-class reliability and superior performance over time. The flat-top beam profile of the solid-state lasers minimizes the effects of changes in fluidics or optics, which in turn can lead to instability or alignment issues and instrument downtime.

 

Laser misalignment is a major concern with users of conventional flow cytometers. The flat-top lasers used in the Attune Flow Cytometers have an intensity profile that allows a wider window of alignment over Gaussian lasers used in traditional systems.  The flat-top lasers also have a higher tolerance for misalignment that allows them to maintain high sensitivity and low CVs.

Emission profile of lasers

 

Gaussian laser used in traditional cytometers (left) with misalignment and flat-top laser used in the Attune Flow Cytometers (right) showing proper alignment if shifted.


Rare event detection

Detection of rare events requires acquisition of high numbers of cells to attain a reliable measure of accuracy. Attune Flow Cytometers allow dilute samples to be processed quickly at sample input speeds of up to 1 mL/min, significantly faster than conventional cytometers that support maximum sample input rates of 60–100 µL/min. Acoustic focusing thus offers a unique combination of speed and quality, cutting the time to collect rare events significantly over long acquisition times.

 

 

Collecting more than 1 million live cells and detecting a rare population of dendritic cells

 

Plasmacytoid dendritic cells (pDCs) are identified using immunophenotype CD19–/B220high/CD317+. Four-color staining of mouse splenocytes included CD19-Pacific Blue, CD317-Alexa Fluor 488, CD45R/B220-PE direct conjugates, and SYTOX AADvanced Dead Cell Stain. A gate was made on live cells using SYTOX AADvanced Dead Cell Stain, followed by gating on CD19– cells. A two-parameter plot of CD45R/B220 vs. CD317 was used to identify pDCs. A collection rate of 500 μL/min was used to acquire 1.3 million total cells with a cell concentration of 7.5 x 107 cells/mL. Plasmacytoid dendritic cells were identified as dual B220+/CD317+ (upper right quadrant) and constitute 0.851% of live CD19– cells, which is 0.194% of total splenocytes.

Attune Flow Cytometers support both high speed of acquisition and high sensitivity to enable easy detection and phenotypic and functional characterization of rare cells. This step-by-step strategy for detecting rare events in our Flow Cytometry Learning Center will help you think through the best approach for pre-analytical, analytical, and data analysis phases of your research.


See what our customers have to say

“When all is said and done, it outperforms [the rest] in that it can analyze 35,000 cells per second with acoustic focusing technology. Its accuracy is not diminished even if you conduct analysis at a high flow rate of 100–1,000 µL/min, which drastically shortens the time required for an experiment.”

Dr. Takashi Satoh,

IFReC Assistant Professor

Osaka University, Japan

“Traditional antibacterial sensitivity testing can take 1–3 days depending on how difficult the bacteria are to grow, isolate, and then test for sensitivity. The Attune NxT allows us, within 1 hour, to tell whether or not the drug will effectively treat the infection. Within 3 hours, accurately assay the minimum drug concentration needed to inhibit bacterial growth.”

Kieran Mulroney,

Biomedical Sciences Researcher

Harry Perkins Medical Research Institute

University of Western Australia

“One of the problems that everyone is familiar with who works in flow cytometry is clogging. Clogging is a thing of the past with this instrument. You have so many samples that you have to run, you can’t wait between samples to clear everything out.”

Bruno Sainz,

CLIP Investigator

Universidad Autónoma de Madrid

Madrid, Spain

“We needed to have a flow cytometer that would allow us to take samples from tumors that had tended to clog other machines. Therefore the Attune NxT is the flow cytometer of choice for our application. The Attune NxT has a number of characteristics that are critical for our research… program. Not only does it have acoustic focusing, which will allow us to go at higher sample throughput, but it also has a larger flow cell, so when we’re looking at tumor cells and isolating cells from those tumors we don’t have to worry about clogging and having to spend a lot of downtime getting the instrument up and running again.”

Charles Prussak,

PharmD, PhD

Director of the Cell Therapy Translational Laboratory (CTTL)

University of California San Diego

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