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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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Updated: Jan 11, 2026

Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry

Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry

Published on: March 17, 2015

Comparative Analysis of Sample Loop and Counting Bead-Based Methods for Size-Dependent Bias in Flow Cytometry.

Hye Ji Shin^1,2, Subeen Kim³, Minjeong Kwak⁴

¹Biometrology Group, Division of Biomedical Metrology, Korea Research Institute of Standards and Science, 267 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea.

Analytical Chemistry

|November 19, 2025

Summary

This summary is machine-generated.

This study reveals size-dependent bias in flow cytometry particle counting using beads. A sample loop method and an empirical equation offer a more accurate approach for particle number concentration measurements.

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Last Updated: Jan 11, 2026

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Area of Science:

Particle characterization
Analytical chemistry
Biophysics

Background:

Accurate particle number concentration measurements are critical in clinical, environmental, and industrial fields.
Flow cytometry with counting beads is a common method, but can introduce size-dependent bias.
Differences in size between target particles and counting beads can lead to inaccurate concentration data.

Purpose of the Study:

To systematically compare the conventional counting bead-based method with a sample loop-based method for particle concentration measurement.
To evaluate and explain the size-dependent bias observed in flow cytometry.
To develop a reliable method for mitigating bias in particle concentration measurements.

Main Methods:

Comparison of conventional counting bead-based method with a sample loop-based method.
Experimental measurement of particle concentrations.
Simulations based on force balance analysis to investigate bias mechanisms.
Development of an empirical equation for bias prediction and mitigation.

Main Results:

Both methods yielded similar concentrations for beads of comparable size.
Significant discrepancies in concentration were observed when bead and target particle sizes differed substantially.
Simulations indicated that hydrodynamic forces and Brownian motion influence bead movement differently based on size, causing bias.
The sample loop method demonstrated reduced size-dependent bias.

Conclusions:

Size-dependent bias in particle concentration measurements using flow cytometry is attributed to differing hydrodynamic behaviors.
The sample loop method offers a more reliable approach by minimizing this bias.
An empirical equation can predict and mitigate bias, improving the accuracy of particle number concentration measurements.