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Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
Published on: March 17, 2015
Aptasensor-enabled quantitative analysis of nano-sized extracellular vesicles by flow cytometry.
Jing Du1, Chao Yuan, Weijie Wang
1Laboratory of Biomedical Microsystems and Nano Devices, Center for Bionic Sensing and Intelligence, Institute of Bio-medical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China. hui.yang@siat.ac.cn jing.du@siat.ac.cn.
A new fluorescent aptasensor method accurately quantifies nano-sized extracellular vesicles (EVs) using high-resolution flow cytometry. This technique offers a sensitive and convenient tool for analyzing EVs in various biological samples.
Area of Science:
- Biotechnology
- Nanotechnology
- Cell Biology
Background:
- Extracellular vesicles (EVs) are crucial for intercellular communication in health and disease.
- Aptasensors offer high selectivity and sensitivity for EV analysis.
- Fluorescent aptasensors combined with flow cytometry (FCM) provide accurate and convenient EV quantification.
Purpose of the Study:
- To develop a fluorescent aptasensor-based method for high-resolution quantitative analysis of nano-sized EVs.
- To demonstrate the utility of the method for analyzing EVs from different cell sources.
- To explore the potential of the technique for evaluating EV bioactivity.
Main Methods:
- Development of a fluorescent aptasensor targeting CD63 on EVs.
- Utilizing a dual-staining procedure with a cytoplasmic dye.
- Employing high-resolution flow cytometry (FCM) for detection and quantification of EVs as small as 100 nm.
Main Results:
- Successful detection and quantification of nano-sized EVs using the developed fluorescent aptasensor.
- Analysis of EVs from bone marrow mesenchymal stem cells, human neural stem cells, and human cornea epithelial cells.
- Quantified EV amounts ranging from 6.79 × 10^6 mL^-1 to 2.08 × 10^8 mL^-1 in culture media.
Conclusions:
- The developed fluorescent aptasensor-FCM method enables accurate quantification of nano-sized EVs.
- This technique is versatile and applicable to EVs from various biological sources.
- The method shows potential as a valuable tool for EV analysis and bioactivity evaluation in future research.
