Attune NxT flow cytometer

What’s new in Attune NxT Software version 4.2

This version of software is on Windows 10 operating system to address the Windows 7 operating system end-of-life by Microsoft. Windows 10 upgrade kit (Cat. No. A41324) and service installation (Cat. No. A46758) is required prior to installation of the version 4.2 software. Version 4.2 includes the option to purchase option to access 21 CFR part 11 software for security, auditing, and electronic signatures in conjunction with the Applied Biosystems SAE server. Finally, this version of software will enable the CytKick and CytKick Max autosampler in a Windows 10 environment.

Additional features include:

  • Improved DPI Support for high resolution monitors
  • Maintenance reminders for system decontamination and post-acquisition rinse
  • Improved behavior of run protocol settings management of experiment versus sample level settings

What’s new in Attune NxT Software version 3.2.1

This version of software is on Windows 7 operating system and facilitates use of CytKick and CytKick Max autosampler.

Download release notes v3.2.1

The Attune NxT software enables powerful user-defined experimental analysis via an intuitive, easy-to-use interface. Advanced tools provide simple acquisition and analysis for users at all experience levels.

Ease of use

  • Intuitive functionality—adjust flow rates, compensation and voltages through intuitive user interface
  • Learn quickly—simple user interface with guided workflows
  • Easy instrument operation—automated maintenance and instrument performance monitoring

Speed

  • Quickly run flow cytometry experiments; less scrolling and clicking
  • Increase productivity; live-streaming update of statistics during acquisition of events
  • Fast refresh rates for large data sets of up to 20,000,000 events per sample with option to append

Guided functionality

  • Guided functions for automated or easy user-defined compensation
  • Automated maintenance: Start Up, Shutdown, Rinse, Sanitize Attune SIP, Deep Clean, Sanitize, System Decontamination, Autosampler Calibration
  • Automated Data and Database backup to help ensure data redundancy

21 CFR Part 11 Compliance*

  • Provides a full audit trail, under which electronic records and electronic signatures are considered trustworthy, reliable, and equivalent to paper records.
  • Authenticates user’s log-in information
  • Locks out access and requires re-authentication
  • Logs any unauthorized attempts
  • Verifies each user’s permissions
  • Permits e-signature function for data verification

*Not included with Attune NxT Software; 21 CFR Part 11 license must be purchased separately.

Visual management buttons and icons help navigate workflows and running experiments. The software allows for:

  • Customizable experimental workflows–pre-define settings for plots, gates, experiments, reports, and overlays
  • Preview screens–easy set-up by showing all plot combinations instantaneously for easy experimental set-up
  • Guided functions–for automated or easy user-defined compensation 

Versatile

  • Heat map for easy setup of plate based assays
  • Sample Recovery to return unused sample to save precious samples
  • Hierarchy view of plots to instantly view complex gating strategies

Customizable

  • User customizable Wait-to-Record function
  • Ability to set user options for default settings for gates, plots, fonts, colors, and group/sample names
  • Visual appearance of the plots is completely customizable; fonts, colors, titles legends and much more can appear exactly the way you want

Publication quality data

  • Smartgate labeling to customize quad gate names and target names
  • Add text, statistics and even images to immediately present data
  • One-click saving of high resolution plots in a variety of file formats including XML, FCS, and even ZIP, for multiple files concurrently
  • Overlay module to perform comparative analysis of single and dual parameter data
  • Preview plots to instantly view all combinations of parameters in a file

User management

  • Levey-Jennings and Performance History reports of baseline and performance tests to monitor trends
  • Ability to create and manage multiple user accounts
  • System access based on user account privileges

System logs amount of time between user logon / logoff and number of tubes run per account per period of time

Note: The Attune NxT Flow Cytometer includes a dongle and 1 license for the Attune NxT Software. The Attune NxT software may be downloaded onto more than one computer, however the software will only run on computers that also have a dongle inserted. Users can also export FCS files in 3.0 and 3.1 format for use in other analysis programs (i.e., FlowJo™ Software).

Software simplifies data acquisition with a dashboard that provides the tools for immediately viewing acquisition status and for setting acquisition stopping criteria. The application settings feature automatically adjusts voltage settings to daily changes in cytometer performance, so that users can run existing applications more simply and quickly.

screenshot showing the Attune software interface
⊕ Enlarge
Intuitive software interface with familiar workflow. The user interface is divided into 4 panels for simple use: (A) the Top panel uses ribbons and tabs for most functions (similar to Microsoft™ Office™ products); (B) the Collection panel presents an easy setup window to acquire data; (C) the Experiment Workspace panel offers display of plots and statistics; and (D) the Experiment Explorer panel makes it easy to manage samples and data files.

Compensation is both rapid and accurate using a guided software system.  Users are only required to prepare the proper controls and adjust the voltage gates.  The system adjusts the compensation and applies it to the samples.

Compensation tools

  • Automated compensation setup allows rapid and accurate compensation and helps to eliminate tedious trial and error adjustments
  • Both negative and unstained gating parameters are available
  • On-plot compensation for fine-tune adjustment
  • Modification of compensation to add or remove parameters as needed after compensation is set up
  • Setup and collect compensation controls directly from a plate
6-step overview of how to apply compensation
(1) Negative gating is useful for heterogeneous samples that are often found in tumor and blood samples. (2) Choose fluorophore-labeled antibodies based on the emitted emission spectrum. Dyes should have minimally overlapping spectra. (3) Prepare single-stained controls. An unstained control is not required. (4) Fluorescent compensation beads can also be used to set compensation parameters when there is not enough sample. (5) Use the guided system and select for negative gate. (6) Run sample and adjust the voltages to see the fluorescent label on the plot. Press Apply Compensation.

Compensation
(1) Unstained compensation is useful for homogeneous samples such as cell lines or already purified cell populations. (2) Chose fluorophore-labeled antibodies based on the emitted emission spectrum. Dyes should have minimally overlapping spectra. (3) Prepare both unstained and single stained controls. (4) Fluorescent compensation beads can also be used to set compensation parameters when there is not enough sample.  (5) Use the guided system and select for unstained gate. (6) Run sample and adjust the voltages to see the fluorescent label on the plot. Press Apply Compensation. 

What’s new in Attune NxT Software version 4.2

This version of software is on Windows 10 operating system to address the Windows 7 operating system end-of-life by Microsoft. Windows 10 upgrade kit (Cat. No. A41324) and service installation (Cat. No. A46758) is required prior to installation of the version 4.2 software. Version 4.2 includes the option to purchase option to access 21 CFR part 11 software for security, auditing, and electronic signatures in conjunction with the Applied Biosystems SAE server. Finally, this version of software will enable the CytKick and CytKick Max autosampler in a Windows 10 environment.

Additional features include:

  • Improved DPI Support for high resolution monitors
  • Maintenance reminders for system decontamination and post-acquisition rinse
  • Improved behavior of run protocol settings management of experiment versus sample level settings

What’s new in Attune NxT Software version 3.2.1

This version of software is on Windows 7 operating system and facilitates use of CytKick and CytKick Max autosampler.

Download release notes v3.2.1

The Attune NxT software enables powerful user-defined experimental analysis via an intuitive, easy-to-use interface. Advanced tools provide simple acquisition and analysis for users at all experience levels.

Ease of use

  • Intuitive functionality—adjust flow rates, compensation and voltages through intuitive user interface
  • Learn quickly—simple user interface with guided workflows
  • Easy instrument operation—automated maintenance and instrument performance monitoring

Speed

  • Quickly run flow cytometry experiments; less scrolling and clicking
  • Increase productivity; live-streaming update of statistics during acquisition of events
  • Fast refresh rates for large data sets of up to 20,000,000 events per sample with option to append

Guided functionality

  • Guided functions for automated or easy user-defined compensation
  • Automated maintenance: Start Up, Shutdown, Rinse, Sanitize Attune SIP, Deep Clean, Sanitize, System Decontamination, Autosampler Calibration
  • Automated Data and Database backup to help ensure data redundancy

21 CFR Part 11 Compliance*

  • Provides a full audit trail, under which electronic records and electronic signatures are considered trustworthy, reliable, and equivalent to paper records.
  • Authenticates user’s log-in information
  • Locks out access and requires re-authentication
  • Logs any unauthorized attempts
  • Verifies each user’s permissions
  • Permits e-signature function for data verification

*Not included with Attune NxT Software; 21 CFR Part 11 license must be purchased separately.

Visual management buttons and icons help navigate workflows and running experiments. The software allows for:

  • Customizable experimental workflows–pre-define settings for plots, gates, experiments, reports, and overlays
  • Preview screens–easy set-up by showing all plot combinations instantaneously for easy experimental set-up
  • Guided functions–for automated or easy user-defined compensation 

Versatile

  • Heat map for easy setup of plate based assays
  • Sample Recovery to return unused sample to save precious samples
  • Hierarchy view of plots to instantly view complex gating strategies

Customizable

  • User customizable Wait-to-Record function
  • Ability to set user options for default settings for gates, plots, fonts, colors, and group/sample names
  • Visual appearance of the plots is completely customizable; fonts, colors, titles legends and much more can appear exactly the way you want

Publication quality data

  • Smartgate labeling to customize quad gate names and target names
  • Add text, statistics and even images to immediately present data
  • One-click saving of high resolution plots in a variety of file formats including XML, FCS, and even ZIP, for multiple files concurrently
  • Overlay module to perform comparative analysis of single and dual parameter data
  • Preview plots to instantly view all combinations of parameters in a file

User management

  • Levey-Jennings and Performance History reports of baseline and performance tests to monitor trends
  • Ability to create and manage multiple user accounts
  • System access based on user account privileges

System logs amount of time between user logon / logoff and number of tubes run per account per period of time

Note: The Attune NxT Flow Cytometer includes a dongle and 1 license for the Attune NxT Software. The Attune NxT software may be downloaded onto more than one computer, however the software will only run on computers that also have a dongle inserted. Users can also export FCS files in 3.0 and 3.1 format for use in other analysis programs (i.e., FlowJo™ Software).

Software simplifies data acquisition with a dashboard that provides the tools for immediately viewing acquisition status and for setting acquisition stopping criteria. The application settings feature automatically adjusts voltage settings to daily changes in cytometer performance, so that users can run existing applications more simply and quickly.

screenshot showing the Attune software interface
⊕ Enlarge
Intuitive software interface with familiar workflow. The user interface is divided into 4 panels for simple use: (A) the Top panel uses ribbons and tabs for most functions (similar to Microsoft™ Office™ products); (B) the Collection panel presents an easy setup window to acquire data; (C) the Experiment Workspace panel offers display of plots and statistics; and (D) the Experiment Explorer panel makes it easy to manage samples and data files.

Compensation is both rapid and accurate using a guided software system.  Users are only required to prepare the proper controls and adjust the voltage gates.  The system adjusts the compensation and applies it to the samples.

Compensation tools

  • Automated compensation setup allows rapid and accurate compensation and helps to eliminate tedious trial and error adjustments
  • Both negative and unstained gating parameters are available
  • On-plot compensation for fine-tune adjustment
  • Modification of compensation to add or remove parameters as needed after compensation is set up
  • Setup and collect compensation controls directly from a plate
6-step overview of how to apply compensation
(1) Negative gating is useful for heterogeneous samples that are often found in tumor and blood samples. (2) Choose fluorophore-labeled antibodies based on the emitted emission spectrum. Dyes should have minimally overlapping spectra. (3) Prepare single-stained controls. An unstained control is not required. (4) Fluorescent compensation beads can also be used to set compensation parameters when there is not enough sample. (5) Use the guided system and select for negative gate. (6) Run sample and adjust the voltages to see the fluorescent label on the plot. Press Apply Compensation.

Compensation
(1) Unstained compensation is useful for homogeneous samples such as cell lines or already purified cell populations. (2) Chose fluorophore-labeled antibodies based on the emitted emission spectrum. Dyes should have minimally overlapping spectra. (3) Prepare both unstained and single stained controls. (4) Fluorescent compensation beads can also be used to set compensation parameters when there is not enough sample.  (5) Use the guided system and select for unstained gate. (6) Run sample and adjust the voltages to see the fluorescent label on the plot. Press Apply Compensation. 

Attune NxT Software releases

Attune NxT Software version 3.1

Thermo Fisher Connect

  • Register an Attune NxT Flow Cytometer with the Thermo Fisher Connect to view performance test data on the cloud.
  • Upload and download experiments and experiment data directly to Thermo Fisher Connect for easy of sharing and data storage.
  • Registered account holders receive 1 TB of free, secure storage built on the world-class Amazon Web Services (AWS) cloud-computing platform.

Hyperlog Transitional slider bars

  • Quickly and easily adjust the HyperLog Transitional value using slider bars on dual-parameter or single-parameter plots.

Easier experiment set up

  • Use Sample Lists or the Sample List view to import/export sample annotation from CSV or Microsoft Excel files.
  • Copy/paste samples in seconds using the heat map view for quick and easy experiment set up that includes the full transfer of instrument settings, workspaces and run protocols.

Gated FCS data

  • Limit file size by recording data within a specific gate, or export data from a specific gate to a new FCS file.

Other improvements

  • Bendable quadrant gates for more precise gating.
  • Increased flexibility for the collection of compensation controls during automated flow cytometry experiments.
  • Drag-n-drop of FCS files with automatic naming for easier import.
  • Improved automated decontamination instructions.

Download v3.1 release notes

Attune NxT Software version 2.7

  • Enhancements to experiment setup and annotation:
    • Import and Export of CSV/Excel files as a Sample List to define groups, samples, experiment notes and custom keywords
    • Copy and paste of cells within the heat map for transfer of run protocols, workspaces and instrument settings
    • Enhanced annotation using a Keyword manager to create user customizable keywords and Sample list view
  • Enhancements to Acquisition controls:
    • Run a plate vertically or horizontally
    • Automatic update of run protocol settings to all samples within an experiment
    • Real-time Acquisition-time Metrics: Elapsed and remaining run time is now displayed on status bar
    • Gated FCS Data: Limit file size by recording of data within a specified gate
    • Improved identification of samples with unique instrument settings using new Instrument Settings badges that label samples with different instrument settings within an experiment
  • Enhancements to data analysis and data processing:
    • Import of individual FCS files by drag-and-drop from the Windows explorer
    • Gated FCS Data: Export of data from a specific gate to a new FCS file
    • Bendable Quadrant gates
    • Copy and paste of the heat map image which summarizes user-processed data to an external application such as Microsoft® Power Point

Download v2.7 release notes

Attune NxT Software version 2.6

In addition to features already incorporated into the Attune NxT software such as auto-compensation, sample recovery, overlays, automated database backup and automated maintenance scripts, our latest software release includes:

  • Improvements to plate-based acquisition using the Attune NxT Autosampler:
    • Pause/Resume for a plate-based acquisition–easily pause acquisition and resume when you’re ready
    • Robotics compatibility–Incorporate the Thermo Scientific Orbitor RS 2 Microplate Mover and SmartStor temperature control device for a comprehensive multi-component automated flow cytometry workcell to maximize operating capacity, mitigate human operator error and enable reproducible data at a high volume. thermofisher.com/flowautomation
  • More control of Experiment Template files–now delete and managetemplates using the improved template manager dialogue.
  • More user-feedback during for sample collection–clear reporting of lost events due to coincidence during acquisition in the “Abort Count” section of the collection panel:

“Exclude Coincidence” OFF

ex-co-off

“Exclude Coincidence” ON

ex-co-on
  • Greater Flexibility for Workspaces: now turn off back-gating

version 2.5 and earlier software (auto-backgate)

attune-2-6

version 2.6 software (customizable-backgate)

attune-2-5
  • Support of new configurations of the Attune NxT Flow Cytometer.  The Attune NxT now includes options for 1-4 laser, including configurations that have 6 detectors for the violet (405 nm) laser. Visit www.thermofisher/attune for more information.

Example 4 laser system with 4 violet channels

filters1

Example 4 laser system with 6 violet channels

filters2

Attune NxT Software version 2.5

In addition to features already incorporated into the Attune NxT software such as auto-compensation, user and configuration-management, sample recovery and automated maintenance scripts, our latest software release includes:

  • Overlay Module: allows easy comparison of results between different samples or between different channels using our overlay module. Users can rapidly create histograms or dot plots comparisons using our fully customizable overlay builder. Great for publications!
attune-img1
  • Peace of mind using our automatic Database Utility program: regularly backup user data with ease using our database utility program. Data is preserved in a user-defined location at a schedule and frequency that meets your needs.
attune-img2
  • Worried about bubbles? Use our automated Bubble Detection System, active for acquisition from both tube and plate samples. Users are notified if a bubble is detected by the system and given the option to recover sample using our unique Sample Recovery option.
  • Fluorescence Compensation now made easier: take advantage of our auto-compensation module and apply compensation recorded from tube or plate samples. Don’t forget our software includes post-acquisition compensation tools that allow the user additional options for accurate data analysis.
  • Improvement to flexible acquisition from 96 and 384 well plates: take advantage of improvements to our plate-mode acquisition and use a ‘manual well’ to optimize and adjust instrument settings directly from the plate. When finished you may record samples in any order needed using customizable options for acquisition from a plate.
  • More control for sample collection: Need to delay how long after a sample is aspirated before it is recorded from a plate? No problem using our new software; improvements to our collection panel allow the user to ‘Wait Before Recording’.
  • Other improvements integrated in this software release include printing, automatic calibration of the Attune NxT Autosampler, improved labeling of channels and compensation controls for easier identification of fluorophore channels in addition to a significant number of bug fixes.

Download v2.5 release notes

Attune NxT Software version 2.4

New features:

  • Support for deep well plates up to 2 mLs
  • Sample recovery to return unused sample to a tube or preloaded sample in a plate
  • Drag and Drop of Experiment Explorer Attributes (Instrument Settings, Workspace, Compensation)
  • Filter Configuration for system wide annotation of fluorophores
  • Implemented context menus on the heat map
  • Completion of optional FCS file keywords (ie $EXP, $INST, $OP, $CELLS, etc.)

Download v2.4 release notes

Attune NxT Software version 2.3

New features:

  • Area Scaling Factors are now applied from Performance Test (Advanced/System Settings)
  • Import and Export of Workspaces
  • Import and Export of Compensation Settings
  • Multi-draw support for deep well plates
  • User options to set Plot label display name ($PnN vs. $PnS)
  • Implemented load instrument settings and workspace on New Experiment dialog
  • Support for Blue, Red, Yellow (BRY) instruments 

Download v2.3 release notes

Attune NxT Software version 2.2

New features:

  • Concentration statistic–obtain concentration measurement from acquired samples
  • Single User Templates–experimental templates available within a user profile
  • Export of FCS files in FCS 3.0 format–in addition to export of FCS 3.1
  • Heat Map Analysis–visual indicator of key results across a plate experiment
  • Import of instrument settings from FCS
  • Statistics Default Options–defined in user profile options
  • Export and Print of the Results Table–for your records

Fixes:

  • Ability to delete multiple plates at the same time using the experiment explorer
  • Ability to multi-select wells and tubes on the experiment explorer
  • Display of the quadrant gates in the edit gates menu
  • Ability to set stop options using quadrant and derived gates

Download v2.2 release notes

Attune NxT Software version 2.1

New features:

  • On-plot compensation
  • Export of statistics to CSV
  • Support of all optional and Attune custom FCS keywords
  • Ability to create and manage multiple user accounts (individual password security)
  • System access based on user account privileges
  • Levey-Jennings and Performance History reports of baseline and performance test
  • Target and Label Fields now carry over to axis labels and can be applied Smart Gate Labeling
  • Append and Overwrite of FCS files
  • Ability to move gate labels (except quad gates)
  • Export and Copy and Paste of plots to external programs (like Power Point)
  • Ability to set user options for default settings for gates, plots, fonts, colors, and group/ sample names

New autosampler features:

  • Support of 384 well plates
  • Support of Deep Well plates—up to 800 µL draw volumes (no multi-draw)
  • Lower Dead Volume—30 µL from wells (for sample flow rates of 12.5 to 500 µL/min)

Download v2.1 release notes

Attune NxT Software version 2.0

New features:

Automated Instrument Functions:

  • Automated Maintenance: Start Up, Shutdown, Rinse, Sanitize Attune SIP, Deep Clean, Sanitize, System Decontamination, Autosampler Calibration

Sample Acquisition:

  • Sample acquisition from flow cytometry tubes, microfuge tubes, or standard 96 well plates using 12.5 μL/min, 25 μL/min, 100 μL/min, 200 μL/min, 500 μL/min, or 1000 μL/min flow rates.
  • Use of round bottom plates is recommended for optimal mixing. Instrument Control functions: Run, Record, Pause, Stop, Clear
  • Creation of plate experiments using the plate set up (“Heat Map”view):
    • Create multiple groups and/or samples within a plate (single experiment per plate only)
    • Create as many individual run protocols as desired per plate \There is a maximum number of 3 mixes recommended at this time
    • Collect samples from the entire plate or only a subset of the plate

Data Visualization Tools:

  • Workspace flexibility: create different workspaces for samples, groups, or an entire experiment
  • Experiment Workspace for easy setup including:
    • Histogram Plot, Dot Plot, Density Plot, Precedence Density Plot; Histogram marker, Rectangle Gate, Polygon Gate, Quadrant Gate and Derived Gates (customized logic gates)
  • Experiment Workspace Layouts: Auto layout or Freeform Workspace options
    • Plot resize tools: use to make plots identical sizes in Freeform mode
    • Plot organization tools: align left or right in Freeform mode
    • Copy and paste gates between plots; change gate type after creation
    • Unfilled or opaque gates

Statistical Measurements Available:

  • Count, % Total, % Gated; Mean, Median, Mode; Standard Deviation, Robust Standard Deviation; % CV, Robust % CV

Viewing Options:

  • “Zoom In” tool that uses a “filmstrip” for easy navigation between plots;
  • Display all events or only a subset of events
  • Customizable Application Layout: move and dock the Collection, Customize, Instrument Settings and Experiment Explorer panels where you would like
  • FCS information viewer: quick access to information on individual FCS file
  • Compatible with multiple monitor displays (note that a single monitor is included with a base package of the Attune® NxT Acoustic Cytometer).

Compensation:

  • Auto-compensation set up and auto-set up of compensation workspaces
  • Acquisition of compensation controls from tubes
  • Background fluorescence set using a negative gate or unstained control
  • Auto-calculation of compensation 

Download v2.0 release notes

Attune NxT Software version 1.1

In addition to features already incorporated into the Attune NxT software such as auto-compensation, user and configuration-management, and automated maintenance scripts, our latest software release includes the following fixes:

  • Baseline and Performance Test updates
  • Hyperlink to set populations available on the top of the plot
  • Updates to instrument settings panel
  • Gate on compensation scatter plot can be applied to all compensation tubes
  • Compensation control is highlighted in voltage adjustment panel
  • Collection panel is updating progress as file is progressing toward stop criteria
  • Number of events displayed is the number selected in stop criteria
  • Changing manual scale can be performed during acquisition
  • Changing plots during acquisition on large event files
  • Copy/Paste of a plot with a gate
  • Key words updated
  • Decontamination Script updates

Download v1.1 release notes

Attune NxT Software version 1.0

The Attune NxT software version 1.0 includes the following features:

Instrument Controls

  • Automated Performance Testing- Baseline and PT (Report)
  • Automated Maintenance- Start up, Shutdown, Rinse, Deep Clean, Sanitize, Decon
  • Data Collection from tubes

Visualization Tools

  • Dot Plot, Density Plot, Histogram
  • Histogram marker, quad marker, Precedence Density Plots (global)
  • Customized logic gates
  • Data Display-Log, Linear
  • HyperLog transform

Statistics

  • Filmstrip Zoom Tool
  • Plot Previews function for easy incorporation of ready-made plots

Experiment Set Up

  • Auto-compensation from tubes
  • Auto layout and Freeform Workspace Layouts

File Management

  • Import/ Export of Files and experiments
  • Files Stored as FCS3.1 format

User Management & Security

  • 3 accounts – single service, admin and user account

Download v1.0 release notes