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Updated: Jun 6, 2026
Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
Published on: March 17, 2015
Flow cytometric analysis of microparticles
Henri C van der Heyde1, Irene Gramaglia, Valéry Combes
1Cell Analysis Core Facility, Flow Cytometry, La Jolla Infectious Disease Institute, San Diego, CA, USA.
Abstract:
Cell-derived microparticles (MPs) are increasingly recognized as important cell-to-cell signaling mechanisms and may exhibit important functions in homeostasis but also in pathogenesis. Indeed, MPs are associated with a number of diseases inhibiting their production that protects against pathogenesis. MPs are distinct from exosomes and apoptotic bodies, often exhibiting the membrane proteins of the activated or apoptotic cell from which they are derived. Electron microscopic analyses have shown that MPs are produced by all cell types tested to date, and ELISA-based assays have established that increased numbers of MPs are produced following cell activation. These approaches do not, however, determine the exact number of MPs and distribution of functional proteins on their surface. Flow cytometry represents an obvious approach to analyze MPs, and we present here a method to assess the number and phenotype of MPs by using a conventional flow cytometer. We also present the caveats with this method and describe a new imaging flow cytometry approach that overcomes these limitations.
Insights
Cell-derived microparticles (MPs) are key for cell signaling and disease. This study presents flow cytometry methods to quantify MP numbers and surface proteins, improving disease diagnostics.
Area of Science:
- Cell Biology
- Immunology
- Biotechnology
Background:
- Cell-derived microparticles (MPs) are vital for intercellular communication in health and disease.
- MPs, distinct from exosomes and apoptotic bodies, carry surface proteins from their parent cells.
- Current methods like electron microscopy and ELISA lack precision in quantifying MPs and their surface markers.
Purpose of the Study:
- To develop and validate a flow cytometry-based method for assessing MP number and phenotype.
- To address limitations of conventional flow cytometry in MP analysis.
- To introduce an advanced imaging flow cytometry approach for comprehensive MP characterization.
Main Methods:
- Utilized conventional flow cytometry to quantify microparticle (MP) numbers and analyze their surface protein expression.
- Identified and described limitations associated with standard flow cytometry for MP analysis.
- Developed and presented an imaging flow cytometry technique to overcome these limitations.
Main Results:
- Established a flow cytometry protocol for enumerating MPs and determining their phenotype.
- Highlighted critical caveats of conventional flow cytometry in MP quantification and characterization.
- Demonstrated the efficacy of imaging flow cytometry in providing a more detailed analysis of MPs.
Conclusions:
- Flow cytometry offers a viable approach for analyzing cell-derived microparticles (MPs).
- Imaging flow cytometry provides enhanced capabilities for precise MP quantification and phenotyping.
- These advancements facilitate a deeper understanding of MP roles in physiological and pathological processes.
