Extracellular vesicles(EVs)are small,membrane-bound structures released by various cell types into the extracellular environment,which play a crucial role in intercellular communication and the transfer of biomolecule...Extracellular vesicles(EVs)are small,membrane-bound structures released by various cell types into the extracellular environment,which play a crucial role in intercellular communication and the transfer of biomolecules between cells.Given their functional signiffcance,there are intense research interests to use EVs as disease markers and drug carriers.However,EVs characterization is greatly hindered by the small size,the low biomolecule payload,and the high level of heterogeneity.To address these challenges,researchers have adopted sensitive microscopic methods such as single-molecule ffuorescence imaging,single-particle dark-ffeld imaging,surface-enhanced Raman scattering,and surface plasmon resonance imaging for single EV analysis.These techniques can detect signals from individual EVs,enabling a detailed study of the heterogeneity.Analysis of EVs cargo has provided insights into the protein/nucleic acid expression and enabled subgroup differ-entiation.Superresolution mapping has visualized EVs structures,and single EV tracking has offered insights into their release and uptake mechanisms.In this review,we will summarize the recent advances in optical imaging of single EVs,including the biomarkers used for EV labeling,the performance of the reported microscopic methods,and their biological ffndings.Finally,we will address the limitationsof the existing methods and outline prospects for future development in this ffeld.展开更多
基金support from the Excellent Young Scientists Fund from the Nation Natural Science Foundation of China and the Zhejiang Leading Innovation and Entrepreneurship Team(2022R01006)the Public Welfare Fund(YXD23H0301)from Natural Science Foundation of Zhejiang,China+1 种基金support from the National Natural Science Foundation of China(Nos.21735006,21890742,22104131)the National Key Scientifc Program of China(2022YFA1304500,2022YFA09133000).
文摘Extracellular vesicles(EVs)are small,membrane-bound structures released by various cell types into the extracellular environment,which play a crucial role in intercellular communication and the transfer of biomolecules between cells.Given their functional signiffcance,there are intense research interests to use EVs as disease markers and drug carriers.However,EVs characterization is greatly hindered by the small size,the low biomolecule payload,and the high level of heterogeneity.To address these challenges,researchers have adopted sensitive microscopic methods such as single-molecule ffuorescence imaging,single-particle dark-ffeld imaging,surface-enhanced Raman scattering,and surface plasmon resonance imaging for single EV analysis.These techniques can detect signals from individual EVs,enabling a detailed study of the heterogeneity.Analysis of EVs cargo has provided insights into the protein/nucleic acid expression and enabled subgroup differ-entiation.Superresolution mapping has visualized EVs structures,and single EV tracking has offered insights into their release and uptake mechanisms.In this review,we will summarize the recent advances in optical imaging of single EVs,including the biomarkers used for EV labeling,the performance of the reported microscopic methods,and their biological ffndings.Finally,we will address the limitationsof the existing methods and outline prospects for future development in this ffeld.
