Attune Flow Cytometer Software

Our suite of image data analysis tools is the main feature of the Attune Cytometric Software version 7.1. We provide workflow wizards that help users with many levels of experience to access the AI machine learning tools. The Image processing feature is intuitive to use and can be incorporated into user-defined scenarios. You can choose a vendor-supplied model that is pre-trained on leukocytes and beads or refine a model to your specifications using the Train Model feature. The system can process images at speeds up to 1,000 images/second, depending on the size and complexity of images. Image-derived data can be exported into the FCS files. Our Model Explorer allows you to visualize stored models, export to share, and import for later use.

Screenshot of new features in the Attune Cytometric Software interface.

Core capabilities

• Compensation—fully automated and manual compensation
• Baseline Functional Response (BFR)—automates the detector optimization process
• Instrument tracking—automated baseline and performance test for all channels and linearity
• Traceability—21 CFR part 11 option for flow cytometry data security, auditing, and electronic signature capabilities
• File formats—FCS 3.0 and FCS 3.1

Maintenance

• Automated maintenance—startup (<15 min), shutdown, rinse, wash, debubble, deep clean, monthly decontamination
• User account maintenance—administrative and individual accounts with user log

Visualization

• Image gallery view of imaged events
• Back-gating of images onto flow plots
• Visualize images from specific gate
• Gates—standard, customizable, and derived gates
• Plot previews—easy workspace setup
• Zoom filmstrip viewer—easy visualization and navigation of plots to set gates and plot attributes
• Overlay plot builder
• Heatmap view with color-coding and statistics 
• Graphics resolution—publication-quality images (customizable)

Image processing with ACS 7.1 & Attune CytPix flow cytometer

• Automated image processing using 36 morphology parameters
• Choice of 3 processing pre-trained models and unlimited user trained models with Train Model Wizard
• Correlate morphology with standard flow cytometric fluorescence and scatter parameters

Data analysis with ACS 7.1 & Attune CytPix flow cytometer

• Dimensionality reduction wizard that integrates morphology and fluorescence data
• Image similarity calculations to quickly find events of similar morphology
• Down Sampling and Concatenation wizard to manage and merge large datasets efficiently

Compensation for spectral overlap between fluorescence channels is both rapid and accurate using a guided software system. The software streamlines the compensation process by performing automated voltration (Baseline Functional Response) with Attune Performance Tracking beads, providing starting points for voltages that can still be adjusted if desired. The system adjusts the compensation and applies it to the samples so that you only need to prepare the proper controls and adjust the voltage gates.

Automated compensation supports both negative and universal negative (unstained) controls. The workflow also allows you to set up and collect compensation controls directly from a plate if needed. This automation eliminates trial and error and delivers compensation rapidly and accurately. In addition, on-plot compensation adjustment allows for fine tuning, and compensation can be subsequently modified.

How to use negative gating 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.

One of the key barriers to increased adoption of flow cytometry is its inherent complication—the myriad of instrument settings users must expertly adjust before acquiring data. To obtain high-quality fluorescent flow cytometer data, well-optimized instrument settings are required. Baseline Functional Response automatically sets the optimal voltage for each detector, enabling users to optimize their experiments quickly by suggesting the minimum voltage required for the best signal.