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Isolation and Characterization of Microvesicles from Peripheral Blood
Published on: January 6, 2017
Isolation and Characterization of Circulating Microparticles by Flow Cytometry
1Kidney Research Centre, Ottawa Hospital Research Institute, University of Ottawa, 451 Smyth Rd., Ottawa, ON, Canada, K1H 8M5. dburger@uottawa.ca.
Abstract:
Microparticles are small fragments (0.1-1.0 μm) of cellular membrane which are shed by cells under conditions of stress. As levels of circulating microparticles are elevated in disease, there has been significant interest in their assessment and quantification under pathological conditions. Here we describe a protocol for the isolation of microparticles from plasma samples and their characterization/quantification by flow cytometry. This assay has been employed for the assessment of microparticle levels in both human and animal plasma and may also be modified for the characterization of microparticles from culture media or from other biological samples (i.e., urine).
Insights
This study details a new protocol for isolating and quantifying microparticles, which are cell membrane fragments elevated in disease. The method uses flow cytometry for assessing microparticle levels in plasma and other biological samples.
Area of Science:
- Biochemistry
- Cell Biology
- Medical Diagnostics
Background:
- Microparticles are cell membrane fragments (0.1-1.0 μm) released by stressed cells.
- Elevated circulating microparticle levels are associated with various pathological conditions.
- Accurate assessment and quantification are crucial for understanding disease states.
Purpose of the Study:
- To describe a reliable protocol for isolating microparticles from plasma.
- To detail a method for characterizing and quantifying microparticles using flow cytometry.
- To provide an adaptable assay for diverse biological samples.
Main Methods:
- Isolation of microparticles from plasma samples.
- Characterization and quantification via flow cytometry.
- Adaptation of the assay for cell culture media, urine, and other biological fluids.
Main Results:
- A standardized protocol for microparticle isolation and quantification was established.
- Flow cytometry enables precise characterization of microparticle populations.
- The assay demonstrated applicability across human and animal plasma samples.
Conclusions:
- The described flow cytometry protocol offers a robust method for microparticle assessment.
- This assay is valuable for research in disease states associated with microparticle changes.
- The protocol's adaptability supports broad applications in biological and clinical research.
