The new method promises to enable precise measurement and fluorescence imaging of extracellular vesicles

On May 4, the team of Yang Hui, a researcher at the Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, published the latest research results in the comprehensive academic journal of chemistry “China Chemical Letters”. The scientific research team proposed a new strategy for fluorescent labeling of extracellular vesicles based on micro-nano fluidic platform to achieve accurate measurement and fluorescence imaging of extracellular vesicles.

In this study, researcher Yang Hui of Shenzhen Advanced Hospital Medical Engineering Institute is the corresponding author of the paper, and doctoral students Hu Shi and Hao Rui are the co-first authors of the paper. Shenzhen Advanced Institute is the first unit of thesis.

Extracellular vesicles (EVs) are nanoscale vesicles with membrane structures actively released by cells, carrying bioactive molecules such as nucleic acids, proteins, and lipids, thereby enabling material exchange between cells. However, for a long time, due to its size characteristics in the 100-nanometer range, it has been difficult to achieve reliable visual fluorescence detection, which has limited the in-depth study of EVs.

At present, the commonly used fluorescent labeling methods for EVs mainly include: membrane protein fluorescent probe labeling and lipophilic fluorescent dye labeling. The fluorescent protein probes have large volume, large steric hindrance, and low abundance and strong heterogeneity of EVs membrane proteins, which greatly limits the efficiency of fluorescent labeling. Lipophilic fluorescent dyes have the advantages of simple operation, high fluorescence intensity and small molecular size, and are suitable for efficient labeling and downstream detection of EVs. However, for a long time, limited by the physicochemical properties of lipophilic fluorescent dyes themselves, dye molecules are prone to agglomeration at higher concentrations, forming nanoparticles similar in size to extracellular vesicles, resulting in false positive signals, resulting in inaccurate detection results and measurement deviations for EVs.

In order to solve the above problems, the research team proposed to use micro-nano fluidic technology to increase the fluidity of EVs membranes by rapid and efficient processing of EVs, and achieve efficient fluorescent labeling at very low dye concentrations. Compared with the “gold standard” of the current experimental operation process, this method greatly improves the fluorescence labeling efficiency of external vesicles under the condition of reducing the dye concentration by several times, so as to achieve efficient flow cytometry detection of EVs and intracellular fluorescence imaging.

Schematic diagram of microfluidic platform labeling extracellular vesicles Courtesy of the scientific research team

EVs fluorescent labeling technology is one of the current research hotspots, which is of great significance for the analysis of the biological function of EVs. The high-efficiency fluorescent labeling method for EVs based on micro-nano fluidic platform proposed in this study provides new ideas and technical support for EVs research. Yang Hui said that this method is expected to promote the development of accurate measurement and fluorescence imaging technology of EVs in the future, and become a powerful and universal technology platform. (Source: China Science News, Diao Wenhui)

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