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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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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Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
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Bead-based multiplexed analysis of analytes by flow cytometry.

Henri C van der Heyde1, Irene Gramaglia

  • 1Cell Analysis Core Facility, Flow Cytometry, La Jolla Infectious Disease Institute, San Diego, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 1, 2010
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Summary

This study introduces fluorescent microspheres for high-throughput protein analysis, enabling hundreds of mini Enzyme-Linked Immunosorbent Assays (ELISAs) simultaneously. This novel method overcomes limitations of traditional assays for precise protein quantification and multiplexing.

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Last Updated: Jun 6, 2026

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
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Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Traditional protein analysis methods like Enzyme-Linked Immunosorbent Assay (ELISA) and Western Blotting are limited in throughput and quantification accuracy.
  • ELISA typically analyzes one protein, while Western Blotting analyzes only a few, hindering comprehensive proteomic studies.
  • Western Blotting often provides relative fold differences rather than exact protein quantification.

Purpose of the Study:

  • To develop a novel, high-throughput method for protein analysis using fluorescent microspheres.
  • To enable simultaneous quantification of multiple analytes, overcoming the limitations of current techniques.
  • To reduce assay variability and improve the precision of protein level and state measurements.

Main Methods:

  • Utilized fluorescent microspheres, each uniquely identified by distinct fluorescence in red and far-red channels (10x10 intensity matrix).
  • Coated microspheres with specific capture molecules to perform numerous mini ELISAs.
  • Developed protocols for simultaneous analysis of up to 100 analytes per assay.

Main Results:

  • Demonstrated the capability to perform hundreds of mini ELISAs concurrently on a single platform.
  • Achieved multiplexed analysis of up to 100 different protein analytes.
  • Significantly reduced assay variability compared to conventional methods.

Conclusions:

  • Fluorescent microsphere technology offers a powerful, high-throughput alternative for protein analysis.
  • This method allows for precise quantification and simultaneous measurement of numerous analytes.
  • The developed protocol enhances efficiency and accuracy in proteomic research and diagnostics.