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Flow Cytometry

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: Mar 27, 2026

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Published on: March 5, 2019

Quantitative Flow Cytometry Measurements in Antibodies Bound per Cell Based on a CD4 Reference.

Lili Wang¹, Heba Degheidy², Fatima Abbasi²

¹National Institute of Standards and Technology (NIST), Gaithersburg, Maryland.

Current Protocols in Cytometry

|January 9, 2016

Summary

This study introduces a standardized method for quantifying biomarkers using multicolor flow cytometry. The new procedure calibrates results in antibody-bound-per-cell (ABC) units for improved clinical relevance and instrument independence.

Keywords:

CD4+ lymphocytes antibody bound per cell (ABC)equivalent number of reference fluorophores (ERF)fluorescence calibration multicolor flow cytometry

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Published on: January 7, 2019

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Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity

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Last Updated: Mar 27, 2026

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Discrimination of Seven Immune Cell Subsets by Two-fluorochrome Flow Cytometry

Published on: March 5, 2019

Isolation of CD4+ T-cells and Analysis of Circulating T-follicular Helper cTfh Cell Subsets from Peripheral Blood Using 6-color Flow Cytometry

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Published on: January 7, 2019

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Published on: December 17, 2013

Area of Science:

Clinical laboratory diagnostics
Immunophenotyping
Biomarker quantification

Background:

Multicolor flow cytometry is vital for measuring cell numbers and receptor expression, serving as disease biomarkers.
Current methods lack standardization, hindering clinical utility and instrument independence.

Purpose of the Study:

To detail a procedure for quantifying surface and intracellular biomarkers using flow cytometry.
To establish a standardized method for biomarker measurement in antibody-bound-per-cell (ABC) units.

Main Methods:

Flow cytometer quality control.
Fluorescence intensity calibration with microspheres.
Compensation for fluorescence spillover.
Application of a biological reference calibrator for ABC scale establishment.

Main Results:

A detailed protocol for calibrating multicolor flow cytometer output.
Quantification of biomarkers in antibody-bound-per-cell (ABC) units.
Foundation for instrument-independent biomarker measurements.

Conclusions:

The described procedure enables standardized, quantitative biomarker measurement via flow cytometry.
This approach enhances clinical relevance and facilitates inter-laboratory comparisons.
Ongoing efforts aim for universal instrument and reagent independence in biomarker quantification.