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Updated: Feb 8, 2026

Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
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Analyzing Platelet Subpopulations by Multi-color Flow Cytometry

Published on: June 10, 2025

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Multi-Color FLUOROSPOT Counting Using ImmunoSpot® Fluoro-X™ Suite

Zoltán Megyesi1, Paul V Lehmann1, Alexey Y Karulin2

  • 1Cellular Technology Ltd., Shaker Heights, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 30, 2018
PubMed

Multi-color FLUOROSPOT assays for simultaneous detection of several T-cell cytokines and/or classes/sub-classes of immunoglobulins secreted by B cells have recently become a major new avenue of development of ELISPOT technology. Advances in assay techniques and the availability of commercial test kits stimulated development of multi-color FLUOROSPOT data analysis platforms. The ImmunoSpot® Fluoro-X™ Software Suite was developed by CTL as an integrated data acquisition, analysis, and management solution for automated high-throughput processing of multi-color T- and B-cell FLUOROSPOT assay plates. The Fluoro-X™ software counting module is based on SmartSpot™/AutoGate™ technologies and utilizes CTL's Center of Mass Distance algorithm for the detection of multi-color spots. The Fluoro-X™ software provides an objective, user error-free means for analyzing multi-color FLUOROSPOT data. An integrated quality control module, with optional GLP and CFR Part 11 compliant package and role-based security, enables data validation, review, and approval with complete audit trails. The extensive multi-format data output and presentation capabilities of the Fluoro-X™ software allow further analysis of FLUOROSPOT data using any commercial flow cytometry software and facilitate the generation of professional reports and presentation. In this article, we present a detailed step-by-step workflow for the analysis of a human four-color IFN-γ, IL-2, TNF-α, and GzB antigen-specific T-cell assay using the Fluoro-X Software Suite.

Keywords:
AntibodiesAutoGate™B CellCenter of massCytokinesELISPOTFLUOROSPOTFluorescenceFluoro-X™ImmunoglobulinsLabelMulti-ColorMultiplexObjectiveSmartCount™ Quality ControlSmartSpot™SoftwareSpot countingT cell

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