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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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Related Experiment Video

Updated: Jun 6, 2026

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
11:11

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction

Published on: August 15, 2013

Quantitative fluorescence measurements with multicolor flow cytometry.

Lili Wang1, Adolfas K Gaigalas, Ming Yan

  • 1National Institute of Standards and Technology, Gaithersburg, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 1, 2010
PubMed
Summary
This summary is machine-generated.

This study presents a new method for quantifying protein biomarkers using multicolor flow cytometry. The procedure calibrates flow cytometer output to provide accurate antibody counts per cell for improved clinical diagnostics.

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Last Updated: Jun 6, 2026

Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
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Area of Science:

  • Clinical laboratory science
  • Immunology
  • Biotechnology

Background:

  • Multicolor flow cytometry is vital for clinical diagnostics, including immunophenotyping and disease monitoring.
  • Current quantitative measurement methods lack satisfactory accuracy and consistency.
  • Standardization is needed for reliable biomarker quantification.

Purpose of the Study:

  • To detail a robust procedure for quantifying surface and intracellular protein biomarkers using multicolor flow cytometry.
  • To establish a standardized method for measuring antibodies bound per cell (ABC).
  • To improve the accuracy and reliability of flow cytometry-based biomarker quantification.

Main Methods:

  • Implementing rigorous quality control (QC) for multicolor flow cytometers.
  • Performing fluorescence calibration with microspheres in equivalent number of reference fluorophores (ERF).
  • Applying compensation techniques to correct for fluorescence spillover.
  • Utilizing a biological reference standard to translate ERF to ABC units.

Main Results:

  • A detailed procedure for quantifying protein biomarkers in absolute units (ABC) was established.
  • The method ensures accurate measurement of antibody binding to cells.
  • The calibration process provides a reliable scale for biomarker quantification.

Conclusions:

  • The developed procedure enables accurate and standardized quantification of protein biomarkers via flow cytometry.
  • This approach enhances the reliability of immunophenotyping and disease monitoring.
  • Future efforts aim for instrument and reagent-independent biomarker quantification.