Analyze High Dimensional Single-Cell Functional Data Easily and Rapidly with IsoSpeak

Traditional proteomic data analysis can be complex and time-consuming, creating major obstacles between data generation and meaningful insights. IsoPlexis’ award-winning data analysis software, IsoSpeak, enables researchers to uncover complex functional proteomics data, faster – down to the individual cell, biomarker, and cytokine.

IsoSpeak’s intuitive user interface allows for push-button visualizations, eliminating the need for specialized personnel, extensive training, and tedious analysis required to generate critical results. Once the IsoPlexis instrument run is complete, you can obtain detailed informatics on critical biomarkers–right at your fingertips.

Single-Cell Cytokine Visualizations Within IsoSpeak Software

With IsoSpeak for IsoPlexis applications, researchers can discover key functional differences in populations of heterogeneous cells through analyzing polyfunctionality, the ability of a cell to secrete 2+ cytokines simultaneously. The presence of highly polyfunctional cells has been shown to be predictive of response, survival, pathogenesis and more. IsoSpeak’s toolkit of advanced visualizations empowers researchers to characterize the function of each cell involved in the immune response, revealing key insights from preclinical to clinical discoveries.

With the ability to toggle between conditions, donors, cell markers, cytokines and more, researchers can instantly customize analyses and visualizations to their experimental needs. Below are just some of the powerful visualizations that IsoSpeak has to offer:


3D tSNE visualization generated with IsoSpeak

The t-SNE provides 3D mapping of single-cell functional cytokine data and high dimensional mapping of unique polyfunctional subsets. The t-SNE graph plots cells based on their greatest cytokine functional differences with three different views options:

  1. Donor groups view allows you to stratify data points from your samples by responders and non-responders to therapy.
  2. Polyfunctionality view helps to compare data points by the degree of the polyfunctionality of the samples.
  3. Dominant Functional Group view color-codes cells based on the dominant cytokine profile being secreted.

Polyfunctional Heatmap

Polyfunctional heatmaps can be used to uncover the critical cell subpopulations that exist in the condition/group of interest. Each row of the heat map corresponds to one sample and each column corresponds to a specific single cytokine or combination of cytokines. Comparing the frequency at which various functional and polyfunctional groups are secreted in your samples allows researchers to better investigate differences in cell subpopulations.

Polyfunctional Overview

The single-cell polyfunctional overview visualization reveals the polyfunctionality of your selected samples, calculated as the percentage of single cells secreting two or more cytokines. Elucidate differences in your samples of interest with the ability to filter between donor groups, cell markers, and more.

Polyfunctional Strength Index

The Polyfunctional Strength Index aggregates all single-cell, multi-dimensional secretions from a sample into a single index, making it easy to identify key differences in samples from high-dimensional data. The readout combines the polyfunctionality of a sample (frequency of cells secreting multiple cytokines) with the signal intensities for each single cell across the secreted cytokines of the sample. The displayed Polyfunctional Strength Index is color-coded to show the contributions from different functional categories of cytokines.

Accessing New Layers of Biological Data With IsoSpeak

IsoSpeak’s easy-to-use interface allows for rapid data analysis and the generation of advanced visualizations across our and single-cell applications. The intuitive software enables researchers to easily visualize their data, stratify samples, reveal functional differences, pinpoint biological drivers of response, and more. With a deeper level of proteomic insight and minimal data analysis time and effort, IsoSpeak empowers labs to access critical biomarkers while accelerating the time to meaningful results.

Ready to try IsoSpeak? Take our virtual interactive software tour to see how IsoSpeak software can simplify your data analysis.


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