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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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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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Determination of the Relative Cell Surface and Total Expression of Recombinant Ion Channels Using Flow Cytometry
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Absolute Quantification of Plasma Membrane Receptors Via Quantitative Flow Cytometry.

Yingye Fang1,2, Manasi Malik3, Sarah K England3

  • 1Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO, USA.

Methods in Molecular Biology (Clifton, N.J.)
|April 22, 2022
PubMed
Summary

Quantitative flow cytometry (qFlow) offers precise measurement of plasma membrane receptors. This method, using phycoerythrin (PE) calibration, enables absolute quantification for disease biomarker studies.

Keywords:
BiomarkerPhycoerythrin (PE)Protein quantificationQuantitative flow cytometryRTK

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Area of Science:

  • Cell Biology
  • Biotechnology
  • Immunology

Background:

  • Plasma membrane receptors are key in cellular signaling and disease diagnostics.
  • Measuring receptor abundance is crucial for understanding disease progression.
  • Quantitative flow cytometry (qFlow) is a powerful technique for receptor analysis.

Purpose of the Study:

  • To detail a quantitative flow cytometry (qFlow) protocol for absolute plasma membrane receptor quantification.
  • To establish a standardized method for measuring receptor abundance using phycoerythrin (PE) calibration.
  • To provide a foundational protocol for studies involving tyrosine kinase and G-protein-coupled receptors.

Main Methods:

  • Utilized quantitative flow cytometry (qFlow) with fluorophore-conjugated antibodies.
  • Employed phycoerythrin (PE) as the fluorophore for absolute quantification.
  • Developed a detailed protocol for standardized and reproducible receptor measurements.

Main Results:

  • Achieved absolute quantification of plasma membrane receptors using qFlow and PE calibration.
  • Demonstrated a reproducible method for measuring receptor abundance.
  • Established a foundational protocol applicable to various receptor types and sample sources.

Conclusions:

  • The described qFlow protocol enables accurate absolute quantification of plasma membrane receptors.
  • This method is essential for biomarker discovery and disease diagnostics.
  • The protocol supports ongoing research in receptor quantification for tyrosine kinase and G-protein-coupled receptors.