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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: Mar 31, 2026

Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

Published on: February 12, 2019

High sensitivity flow cytometry of membrane vesicles.

Samuel A Stoner¹, Erika Duggan^1,2, Danilo Condello^1,2

¹La Jolla Bioengineering Institute, La Jolla, California, 92037.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|October 21, 2015

Summary

This study introduces a high-sensitivity flow cytometry method for analyzing extracellular vesicles (EVs). This technique enables precise measurement of EV size, number, and surface markers, advancing their use as diagnostic biomarkers.

Keywords:

extracellular vesicle fluorescence liposome microparticle microvesicle nanoparticle

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Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

Flow Cytometric Analysis of Extracellular Vesicles from Cell-conditioned Media

Published on: February 12, 2019

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

Flow Virometry to Analyze Antigenic Spectra of Virions and Extracellular Vesicles

Flow Virometry to Analyze Antigenic Spectra of Virions and Extracellular Vesicles

Published on: January 25, 2017

Area of Science:

Biotechnology
Cell Biology
Nanotechnology

Background:

Extracellular vesicles (EVs) are crucial for intercellular signaling but their phenotypic heterogeneity poses research challenges.
Conventional flow cytometry struggles to analyze the small size of EVs, limiting their diagnostic and therapeutic potential.
Developing methods to characterize individual EVs is essential for understanding their biological roles.

Purpose of the Study:

To develop and validate a high-sensitivity flow cytometry approach for enumerating and characterizing individual extracellular vesicles (EVs).
To assess the utility of fluorescent membrane probes for accurate EV size and number estimation.
To identify and quantify specific EV subsets using surface marker analysis.

Main Methods:

Construction of a high-sensitivity flow cytometer for EV detection.
Evaluation of fluorescent membrane probes, including di-8-ANEPPS, for vesicle fluorescence proportional to surface area.
Utilizing fluorescence-labeled annexin V and anti-CD61 antibody for surface marker analysis on EVs from rat plasma.
Stimulating platelet-rich plasma with calcium ionophore to observe changes in EV surface marker expression.

Main Results:

The voltage-sensing dye di-8-ANEPPS enabled accurate EV size and number measurements.
Flow cytometry successfully enumerated individual EVs and estimated their size.
Increased fractions of annexin V and CD61-positive EVs were observed after calcium ionophore treatment.
Phenotypic subsets of EVs were identified based on surface marker expression.

Conclusions:

High-sensitivity flow cytometry with fluorescence-based detection is a viable method for analyzing extracellular vesicles (EVs).
This approach allows for the characterization of EV size, number, and surface markers, overcoming limitations of conventional methods.
EV flow cytometry holds significant potential for utilizing cell-derived vesicles as functional biomarkers in various applications.