Attune™ NxT Flow Cytometer, blue

Catalog No.	Quantity
A24864	1 each

Catalog No. A24864 Supplier LIFE TECHNOLOGIES Supplier No. A24864

Description

Attune™ NxT Flow Cytometer, blue

The Attune™ NxT Acoustic Focusing Cytometer is an easy-to-use instrument designed to detect cell size and granularity, as well as fluorescence expression of labeled antibodies, proteins, and functional reagents. The innovative design of the instrument results in precise data and acquisition speeds up to 10x faster than other flow cytometers. The design also eliminates lengthy sample preparation procedures, reduces the time to analyze a file, allows small or limited samples to be analyzed, and minimizes variability in measurements, leading to better resolution of populations. It is ideal for immunophenotyping and signaling studies, cell cycle analysis, detection of rare events, stem cell analysis, cancer and apoptosis studies, and microbiological assays.

With the Attune™ NxT Acoustic Focusing Cytometer you get:

A flexible platform that provides highly sensitive and accurate measurement of cell sizeand granularity, as well as fluorescence detection

State of the art optics for long-term reliability

Acoustic focusing for high-precision data

An intuitive software user interface that reduces time to proficiency

Improved workflow to reduce sample preparation time before analysis

Faster data analysis that leads to reduced acquisition times

Flexible design that accommodates changing needs, budget, or laboratory space
The instrument’s modular design accommodates the most common fluorophores and fluorescent proteins used in flow cytometry enabling alignment with previous experimental results and accommodating future needs through simple upgrades. The small footprint of the instrument requires minimal bench space, even allowing use in a cell culture hood.

Acoustic focusing improves efficiencies compared to traditional flow cytometers
Traditional flow cytometers rely on a large volume of fluid to focus cells prior to analysis, resulting in extensive sample preparation prior to analysis and introducing the possibility of error due to artifacts created during the sample preparation process. With the Attune™ NxT Acoustic Focusing Cytometer, blood samples can be run without being manipulated using the No-Wash No-Lyse blood cell assay. The instrument also offers flexibility in the sample concentration that can be analyzed.

Acoustic focusing enables:

Elimination of concentration steps

Effective background fluorescence reduction by a simple dilution step, eliminating the need to wash fluorescence samples before analysis

No-lyse/no-wash assays

Analysis of samples that have low cell numbers, including CSF, stem cells, mouse bone marrow, mouse blood, and thin needle aspirates

Acquisition of a high volume of samples in minimal time

Learn more about the Attune NxT cytometer and see all available configurations >
Learn more about how acoustic focusing works >

Specifications

• Content And Storage: • Attune NxT Flow Cytometer
  • Minitower Windows 10 64-bit computer with Attune Cytometric Software
  • Power cord with adapters
  • USB cable
  • Attune Shutdown Solution
  • Attune Wash Solution
  • Attune Focusing Fluid
  • Attune NxT Flow Cell Cleaning Solution
  • Attune Performance Tracking Beads
  • Quick reference card
  
  
• Format: Tube(s)
• Laser Features: Blue Excitation Laser (488 nm)
• Frequency: 50/60 Hz
• Product Line: Attune
• For Use With (Equipment): Attune™ NxT Acoustic Focusing Cytometer
• Shipping Condition: Room Temperature
• Quantity: 1 each

Frequently Asked Questions (FAQs)

My Attune NxT Flow Cytometer performance test keeps failing. What steps can I take to minimize the number of failed performance tests?

To minimize the number of failed performance tests on your Attune NxT Flow Cytometer, we suggest the following:

•   Make sure to carry out daily, weekly, and monthly maintenance procedures (pages 8-14 of the Maintenance and Troubleshooting Guide, linked below).
•   Always run 3 startup cycles and 2 Rinse procedures (pages 46-47 of the Maintenance and Troubleshooting Guide linked below).
•   If the performance test fails, carry out the following priming sequence: run 3 startup cycles, 2 de-bubble cycles, and 5 rinse procedures, followed by running the SIP sanitize function.

Please review the Attune Flow Cytometry Maintenance and Troubleshooting Guide for more information about performing routine maintenance procedures and troubleshooting.

Can I use the Attune Nxt Flow Cytometer to detect bacteria?

Yes. The Attune Nxt Flow Cytometer can detect down to 0.2 µm and using a special configuration, down to 0.1 µm. Optimize your settings using Flow Cytometry Sub-micron Particle Size Reference Kit, Cat. No. F13839.

Of all the Attune NxT filters you offer, I don't see one that matches our application. Can I use a filter other than what you offer?

Yes. You would need to purchase Attune NxT Custom Filter Holder Kit, Cat. No. A27784 to make your own custom emission or dichroic filters in order to customize the optical configuration of the instrument. Each kit contains sufficient parts to make 2 emission filters and 2 dichroic filters. The kit includes instructions for attaching the filter to the filter blade, but does not include the glass filter. Other accessories to optimize for special applications include Cat No. 100083194 Attune NxT Small Particle Side-Scatter Filter, Cat. No. 100083194 and Attune NxT Blocker Bar Conversion Kit, Cat No. A35966.

With the Attune Nxt Flow Cytometer, if I wish to only start with a single laser, can I expand to more lasers and channels?

Yes. There are 6 lasers to choose from (blue, green, yellow, red, violet, and violet 6). You can start with one laser and at any time upgrade the same instrument with up to 4 lasers to utilize a maximum of 14 detection channels. Using the Violet 6 laser along with the blue, red, and yellow lasers, one can detect up to 16 channels.

What makes the Attune Nxt Flow Cytometer better than standard flow cytometers?

Because of the acoustic-assisted hydrodynamic technology, the Attune NxT Flow Cytometer uses only 1/10th the amount of fluids, and is 10X faster and less sensitive to clogging compared to standard flow cytometers. You can set the flow rate from 12.5-1,000 µL/min and collect up to 65,000 events/sec using from 20 µL to 4 mL of sample volume. These unique properties allow for rare event analysis, as well standard flow analysis.

How often should I change filters for the Attune NxT Acoustic Focusing Cytometer?

Filters are recommended for replacement once every three months, unless performing small particle detection when once-a-month replacement is recommended.

I've run out of Attune Shutdown Solution for the Attune NxT Flow Cytometer. What can I use for the time being in place of the shutdown solution until I can get more shutdowns solution?

In an emergency, we recommend using the highest purity of water available in place of the shutdown solution and then switching to the Attune Shutdown Solution as soon as possible.

I've run out of Attune Wash Solution for the Attune NxT Flow Cytometer. What can I use for the time being in place of the wash solution until I can get more wash solution?

In an emergency, we recommend using the highest purity of water available in place of the wash solution and then switching to the Attune Wash Solution as soon as possible.

When I export Attune NxT FCS files, do they include the compensation values set in the experiment?

You can tell whether the compensation data are included by looking for the $SPILL or $COMP keyword embedded within the FCS files. You can also see the FCS file information within the Attune NxT software by going to the "View" tab and checking the box labeled "FCS File Panel". This will open a window which shows the information contained in the FCS file associated with the selected sample. It will include any compensation ($SPILLOVER) that has been done.

Is an isotype control a must for flow cytometry analysis?

No. Many users are using unstained cells in combination with FMO controls to identify their positive populations.

I am using the Attune Nxt Acoustic Focusing Cytometer. Why is my event rate so low?

There are several things that can cause a low event rate:

-The system may be clogged
-The threshold level may be set too high
-The PMT voltage for the threshold parameter may be set too low
-The sample may be too dilute or not adequately mixed
-The sample syringe may be loose

I am using the Attune Nxt Acoustic Focusing Cytometer. Why is my event rate so high?

There are several things that can cause a high event rate:

-There may be an air bubble in the flow cell
-The threshold level may be set too low
-The PMT voltage for the threshold parameter may be set too high
-The sample may be too concentrated
-The sample flow rate may be too high
-The sample may have a bacterial contamination

Where can I find the log file for my Attune Nxt instrument?

Open the Attune NxT software, open the main menu, and go to the “Instrument” tab. The option on the far left is “System Log”. Click on that and it will open the log file. At the bottom of the page there is a button labeled “Export”. Use this to export and save the log file.

I ordered a box of Attune Focusing Fluid (10L, A24904) and I cannot find the spigot. What should I do?

We recommend looking inside the box, possibly underneath the bag of buffer.

I am using the Attune Nxt Acoustic Focusing Cytometer and my data does not look like I expected. How can I get help with it?

Send an email to Cellular Analysis Technical Support at technical-molecularprobes@lifetech.com, including the words “flow*#148; or “Attune” in the subject line for proper routing of your email. Please describe the type of application you are doing, how you set up the experiment, and your results. It would be ideal if you could send us a sample of your data that we can review.

I have an Attune Nxt Acoustic Focusing Cytometer and it’s not working right. Whom do I contact?

If the issue is clearly an instrument problem, send an email to Instrument Services at instrumentservices@thermofisher.com with your instrument serial number. If the issue is software or application-related, or if you do not know the source of the problem, send an email to Cellular Analysis Technical Support at technical-molecularprobes@lifetech.com, including the words “flow*#148; or “Attune” in the subject line for proper routing of your email.

Can you help me design my antibody panel for immunophenotyping?

For help with designing panels for flow cytometry, see our new Molecular Probes Flow Cytometry Panel Design Tool (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometry-panel-design-tool.html) The panel design tool can help you choose fluorescent antibody conjugates for your flow cytometry panel in a few easy steps: pick the antibody species reactivity, select up to 14 targets of interest (choices include viability dyes), and choose the lasers or fluorophores you want to view.

How many different parameters can you combine in one assay?

It depends on your instrument. People have done 17 colors (Perfetto SP, Chattopadhyay PK, and Roederer M (2004) Seventeen-colour flow cytometry: unraveling the immune system. Nat Rev Immunol 4:648–655 (http://www.ncbi.nlm.nih.gov/pubmed/15286731)) and more, but it takes a lot of planning and testing to make it work. The Attune NxT Acoustic Focusing Cytometer can run up to 14 colors plus forward and side scatter with a system equipped with all four lasers (violet, blue, yellow and red).

What is an FMO control in flow cytometry?

An FMO control (flow minus one) is a control in which you label cells or beads with every color in your panel, omitting one. Make one FMO control for each color in your panel. These controls are important for helping you properly set gates on your data.

What if I can't find the right isotype control for flow cytomtery?

The flow cytometry field is moving away from using isotype controls as they are not necessarily the most appropriate way to control for non-specific binding. Instead, they are using unstained cells to define the negative population, single stained controls to set compensation, and flow minus one (FMO) controls to set regions and gates.

You may want to consider whether using an isotype control is something you need to do. Here are some references you might want to look at:
-O'Gorman MR, Thomas J. (1999) Isotype controls-time to let go? Cytometry 38:78-80.

-Keeney M, Gratama JW, Chin-Yee IH et al. (1998) Isotype controls in the analysis of lymphocytes and CD34+ stem/progenitor cells by flow cytometry- time to let go! Cytometry 34:280-283.

-Hulspas R, O'Gorman MR, Wood BL et al. (2009) Considerations for the control of background fluorescence in clinical flow cytometry. Cytometry B Clin Cytom 76:355–364.

-Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline - Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H42-A2 Volume 27 No.16, 2007.

-Clinical Flow Cytometric Analysis of Neoplastic Hematolymphoid Cells; Approved Guideline - Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H43-A2 Volume 27 No. 11, 2007.

If you do wish to use isotype controls, we offer a wide variety of species and fluorophores, search our Web site under "isotype antibody control."

What is the Flow Cytometry Size Calibration Kit (Cat. No. F13838) used for?

Our Flow Cytometry Size Calibration Kit has non-fluorescent particle-size calibration standards that provide a simple, accurate way to determine cell sizes by flow cytometry. The kit contains six suspensions of highly uniform polystyrene microspheres with the following diameters: 1.0 µm, 2.0 µm, 4.0 µm, 6.0 µm, 10.0 µm and 15.0 µm. All of the microsphere suspensions are provided in convenient dropper vials. This kit looks at the scatter properties of non-fluorescent microspheres of known sizes to determine approximate cell sizes.

What is the smallest size that I can detect with the Attune NxT Acoustic Focusing Cytometer?

The smallest size that you can detect with the Attune NxT Acoustic Focusing Cytometer is 0.5 µm.

What is the Attune NxT Autosampler?

The Attune NxT Autosampler, an optional accessory for the Attune NxT Acoustic Focusing Cytometer, enables rapid processing of up to 384 samples. It has broad compatibility with different plate formats, both 96- and 384-well plates. It has an intelligent probe designed to minimize clogging and carryover (<0.5%) and to prevent damage to the instrument. It mixes by aspiration rather than shaking to ensure homogeneity of the sample and maintain cell viability. Is performs automated cleaning as part of the shutdown process of the Attune NxT Cytometer. It provides consistent data regardless of sampling method (tube vs. plate) and collection rate.

What are the advantages of acoustic-assisted hydrodynamic focusing in flow cytometry?

-Modular design - Multiple configurations available - field upgradable.
-Save time - 10X faster speeds with no loss in data quality.
-Simplified sample prep - No wash, no lyse options, non-clogging fluidics.
-Enables unique applications - Complex protocols on a broad range of cell types and samples.

How is the Attune NxT Acoustic Focusing Cytometer different from traditional flow cytometers?

With the option to be configured with up to 4 lasers and 14 colors for multi-parameter analysis the Attune NxT Acoustic Focusing Cytometer was designed as a modular system to fit most experimental needs and lab budgets. The novel design of the optical path helps ensure precise fixed alignment of four spatially separated lasers onto the sample stream enabling consistency in data over time, superior performance, and superior reliability. The instrument can be configured with up to 4 solid-state lasers (405 nm, 488 nm, 561 nm, and 637 nm) with flat top beam profiles.

The Attune NxT Flow Cytometer's acoustic focusing uses ultrasonic radiation pressure (> 2 MHz) to transport particles into the center of the sample stream. This pre-focused stream is then injected into the sheath stream, which supplies an additional hydrodynamic pressure to the sample. The combination of these two forces- termed acoustic-assisted hydrodynamic focusing-results in a narrow core stream and uniform laser illumination, regardless of the sample input rate. In traditional cytometers that rely solely on hydrodynamic focusing, the sample core widens to accommodate the increases in flow rate, which results in less uniform laser light illumination.

What is flow cytometry?

Cytometry is the measurement of physical or chemical characteristics of cells or particles. Flow cytometry measures these characteristics of cells or particles as they individually pass lasers in a flow cytometer instrument. Flow cytometry is performed on single cells, providing discrete measurements for each cell in the sample. It also provides a statistical distribution of the measured characteristics of the sample.

A flow cytometer is made up of three subsystems: fluidics, optics, and electronics. Fluidics moves the cells and introduces them for interrogation. Optics generates and collects the light signals. Electronics converts the optical signals to proportional electronic signals for computer analysis.

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