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Acoustic Focusing Cytometry Technology Overview
Novel technology for precision and speed
Equipped with acoustic-assisted hydrodynamic focusing, the Invitrogen Attune Flow Cytometers are able to achieve sample-throughput rates up to 10 times faster than traditional cytometers. This means that you can process samples more quickly and acquire sufficient events even from very dilute samples. The Attune Flow Cytometers accommodate no-wash, no-lyse protocols, allowing you to collect data on samples that have been subjected to minimal handling. In addition, the Attune CytPix Flow Cytometer includes a high-speed, high-resolution, brightfield camera that records images of particles as they pass through the flow cell. The camera and Attune Cytometric Software help ensure that events recorded by the detector are single cells as opposed to doublets, clumps, or debris.
"We have yet to clog the machine with our debris-rich primary tumor samples. Of course, the acoustic technology greatly facilitates the identification of small populations, like cancer stem cells, increasing our capacity to detect and quantify these rare events with high efficiency and reliability."
– Bruno Sainz Jr., PhD, Autonoma University of Madrid, School of Medicine
Acoustic focusing in action
This video demonstrates the event alignment that occurs when acoustic focusing is applied.
Faster sample processing compared to traditional cytometers
Equipped with acoustic-assisted hydrodynamic focusing, the Attune Flow Cytometers process samples faster than traditional cytometers. This breakthrough technology uses sound waves for more precise cell alignment and higher data quality. The pre-alignment of cells allows the Attune Flow Cytometers to process samples quickly and acquire more statistically significant events even from very dilute samples. Acoustic focusing is doubly useful for cellular imaging on the Attune CytPix instrument, helping to position the cells precisely so that a sharp image is obtained.
Acoustic focusing offers many benefits including:
- 10X faster than traditional flow cytometers
- Broadens experimental capabilities by completing time-consuming experiments such as cell cycle analysis faster and without compromising data quality
- Accommodates a broader range cell types, from small platelets to large cardiomyocytes
View from of the end of the acoustic-assisted flow cytometer's capillary with cells run at 500 µL/min.
Principles behind acoustic focusing
Flow cytometry data quality depends on collecting information from thousands of individual cells as they pass through the laser interrogation point. The acoustic technology within Attune Flow Cytometers generate ultrasonic waves that act to transport particles to the center of the sample stream. The pre-focused stream is injected into the sheath stream, which applies hydrodynamic pressure to the sample. The combination of these two forces results in a narrow core stream and an increased probability that cells are passing single file through optical system (Figure 1).
Acoustic-focusing cytometry
Figure 1. Overview of acoustic-assisted hydrodynamic focusing employed in the Attune Flow Cytometers.
Acoustic technology maximizes quality and time
Traditional hydrodynamic focusing permits faster flow rates by broadening the width of the sample stream. This can be problematic as a wider core may allow more than one cell to pass through the laser at the same time (Figure 2B). Acoustic focusing does not broaden the core width to accommodate faster sample flow rates (Figure 2A). This type of system helps preserve data quality as single-cell interrogation is more likely to be maintained even at increased sample flow rates (Figure 2C).
Maximize quality and time
Figure 2. A wider distribution core typically means a broader distribution of cells. (A) Acoustic focusing technology allows for a narrow sample stream regardless of sample input rate. (B) Traditional hydrodynamic focusing requires widening the sample core for faster sample input rates. (C) Instruments equipped with acoustic-assisted technology can accommodate sample input rates from 12.5 to 1,000 µl/min.
Acoustic technology helps the Attune Flow Cytometers achieve sample-throughput rates up to 10 times faster than traditional hydrodynamic focusing systems (Figures 3 and 4) and data acquisition speeds of 35,000 events/sec. Concentrated samples can be diluted and run quickly on the Attune Flow Cytometers, offering your lab rapid high-quality data acquisition capabilities.
Figure 3. Rapid data acquisition. The Attune Flow Cytometers acquires 1,000,000 events in a fraction of the time required compared to three competitor instruments. Data represents protocol time achieved by running the flow cytometers at maximum sample rates for each instrument (1,000 µL/min for the Attune Flow Cytometers).
Figure 4. Minimal data variation at high sample rates with the Attune Flow Cytometers. Jurkat cells were fixed and stained with propidium iodide, treated with RNase and analyzed at a concentration of 1 x 106 cells/mL on the Attune Flow Cytometers. Regardless of sample rate, 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.
Faster acquisition times for the applications you’re already running
We've conducted countless experiments using the Attune Flow Cytometers, many of the same ones scientists are performing in their labs every day. Take a look at the data we've obtained for the following applications:
Analyze more cell types and maintain data integrity
Save time and minimize cell handling with faster protocols
The Attune Flow Cytometers accommodates a no-wash, no-lyse protocol for whole blood, which dramatically reduces the number of steps you have to perform, the amount of sample that is lost during wash steps, and the amount of handling your cells have to endure. See the no wash, no lyse options sample data
Comparison of whole blood immunophenotyping protocols
| Number of steps | Total sample prep time | |
|---|---|---|
| Traditional immunophenotyping protocol | 10 | ~1–2 hr |
| No wash, no lyse immunophenotyping protocol | 3 | ~15–20 min |
Download the application notes:
A Journey Through Innovation, The Discovery of Acoustic Focusing and the Attune Flow Cytometer
This mini-webinar discusses how the discovery of sound measurement lead to a series of discoveries culminating in the knowledge needed to use sound in particle focusing.
Principles behind acoustic focusing
Flow cytometry data quality depends on collecting information from thousands of individual cells as they pass through the laser interrogation point. The acoustic technology within Attune Flow Cytometers generate ultrasonic waves that act to transport particles to the center of the sample stream. The pre-focused stream is injected into the sheath stream, which applies hydrodynamic pressure to the sample. The combination of these two forces results in a narrow core stream and an increased probability that cells are passing single file through optical system (Figure 1).
Acoustic-focusing cytometry
Figure 1. Overview of acoustic-assisted hydrodynamic focusing employed in the Attune Flow Cytometers.
Acoustic technology maximizes quality and time
Traditional hydrodynamic focusing permits faster flow rates by broadening the width of the sample stream. This can be problematic as a wider core may allow more than one cell to pass through the laser at the same time (Figure 2B). Acoustic focusing does not broaden the core width to accommodate faster sample flow rates (Figure 2A). This type of system helps preserve data quality as single-cell interrogation is more likely to be maintained even at increased sample flow rates (Figure 2C).
Maximize quality and time
Figure 2. A wider distribution core typically means a broader distribution of cells. (A) Acoustic focusing technology allows for a narrow sample stream regardless of sample input rate. (B) Traditional hydrodynamic focusing requires widening the sample core for faster sample input rates. (C) Instruments equipped with acoustic-assisted technology can accommodate sample input rates from 12.5 to 1,000 µl/min.
Acoustic technology helps the Attune Flow Cytometers achieve sample-throughput rates up to 10 times faster than traditional hydrodynamic focusing systems (Figures 3 and 4) and data acquisition speeds of 35,000 events/sec. Concentrated samples can be diluted and run quickly on the Attune Flow Cytometers, offering your lab rapid high-quality data acquisition capabilities.
Figure 3. Rapid data acquisition. The Attune Flow Cytometers acquires 1,000,000 events in a fraction of the time required compared to three competitor instruments. Data represents protocol time achieved by running the flow cytometers at maximum sample rates for each instrument (1,000 µL/min for the Attune Flow Cytometers).
Figure 4. Minimal data variation at high sample rates with the Attune Flow Cytometers. Jurkat cells were fixed and stained with propidium iodide, treated with RNase and analyzed at a concentration of 1 x 106 cells/mL on the Attune Flow Cytometers. Regardless of sample rate, 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.
Faster acquisition times for the applications you’re already running
We've conducted countless experiments using the Attune Flow Cytometers, many of the same ones scientists are performing in their labs every day. Take a look at the data we've obtained for the following applications:
Analyze more cell types and maintain data integrity
Save time and minimize cell handling with faster protocols
The Attune Flow Cytometers accommodates a no-wash, no-lyse protocol for whole blood, which dramatically reduces the number of steps you have to perform, the amount of sample that is lost during wash steps, and the amount of handling your cells have to endure. See the no wash, no lyse options sample data
Comparison of whole blood immunophenotyping protocols
| Number of steps | Total sample prep time | |
|---|---|---|
| Traditional immunophenotyping protocol | 10 | ~1–2 hr |
| No wash, no lyse immunophenotyping protocol | 3 | ~15–20 min |
Download the application notes:
A Journey Through Innovation, The Discovery of Acoustic Focusing and the Attune Flow Cytometer
This mini-webinar discusses how the discovery of sound measurement lead to a series of discoveries culminating in the knowledge needed to use sound in particle focusing.
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