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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Related Experiment Video

Updated: Jan 3, 2026

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
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LEGENDplex™: Bead-assisted multiplex cytokine profiling by flow cytometry.

Jason S Lehmann1, Priyanka Rughwani1, Melissa Kolenovic1

  • 1BioLegend, San Diego, CA, United States.

Methods in Enzymology
|November 16, 2019
PubMed
Summary
This summary is machine-generated.

Advances in tumor immunology have yielded new therapies, but understanding the tumor microenvironment is key. This study details bead-based multiplex assays for quantifying critical signaling molecules like cytokines and chemokines.

Keywords:
BiomarkersChemokinesCytokinesLEGENDplex™MultiplexQuantitative immunoassaySoluble mediators

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Area of Science:

  • Immunology
  • Cancer Research
  • Biotechnology

Background:

  • Significant progress in tumor immunology has spurred the development of novel immunotherapy strategies.
  • The tumor microenvironment's complex immune dynamics require further investigation for optimal therapeutic outcomes.
  • Soluble mediators, including cytokines and chemokines, play a crucial role in regulating anti-tumor immunity.

Purpose of the Study:

  • To provide a detailed technical protocol for LEGENDplex™ bead-based multiplex assays.
  • To highlight the applicability of these assays in quantifying critical signaling molecules within the tumor microenvironment.
  • To advance the understanding of tumor immunology and enhance the efficacy of cancer therapeutics.

Main Methods:

  • Detailed description of the LEGENDplex™ assay protocol.
  • Demonstration of multiplex analysis for simultaneous quantification of multiple analytes.
  • Application of the assay for measuring cytokines, chemokines, and other soluble mediators.

Main Results:

  • The study establishes a robust method for quantifying key signaling molecules.
  • The LEGENDplex™ assay demonstrates high sensitivity and specificity for detecting immune mediators.
  • The protocol is applicable to various research settings studying the tumor microenvironment.

Conclusions:

  • LEGENDplex™ bead-based multiplex assays are a valuable tool for studying tumor immunology.
  • Accurate quantification of soluble mediators is essential for understanding immune responses in cancer.
  • This methodology can contribute to the development of more effective cancer immunotherapies.