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单囊泡成像和共定位分析用于个体细胞外囊泡的四跨膜蛋白谱分析。

Single-vesicle imaging and co-localization analysis for tetraspanin profiling of individual extracellular vesicles.

机构信息

Department of Mechanical Engineering Pohang University of Science and Technology Pohang Gyeong-buk Republic of Korea.

School of Interdisciplinary Bioscience and Bioengineering Pohang University of Science and Technology Pohang Gyeong-buk Republic of Korea.

出版信息

J Extracell Vesicles. 2021 Jan;10(3):e12047. doi: 10.1002/jev2.12047. Epub 2021 Jan 11.

DOI:10.1002/jev2.12047
PMID:33456726
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7797949/
Abstract

Extracellular vesicles (EVs) are secreted nano-sized vesicles that contain cellular proteins, lipids, and nucleic acids. Although EVs are expected to be biologically diverse, current analyses cannot adequately characterize this diversity because most are ensemble methods that inevitably average out information from diverse EVs. Here we describe a single vesicle analysis, which directly visualizes marker expressions of individual EVs using a total internal-reflection microscopy and analyzes their co-localization to investigate EV subpopulations. The single-vesicle imaging and co-localization analysis successfully illustrated the diversity of EVs and revealed distinct patterns of tetraspanin expressions. Application of the analysis demonstrated similarities and dissimilarities between the EV fractions that had been acquired from different conventional EV isolation methods. The analysis method developed in this study will provide a new and reliable tool for investigating characteristics of single EVs, and the findings of the analysis might increase understanding of the characteristics of EVs.

摘要

细胞外囊泡（EVs）是一种分泌的纳米大小的囊泡，其中包含细胞蛋白、脂类和核酸。尽管预计 EVs 具有丰富的生物学多样性，但目前的分析方法无法充分描述这种多样性，因为大多数方法都是基于集合的，这不可避免地会平均化来自不同 EV 的信息。在这里，我们描述了一种单细胞囊泡分析方法，该方法使用全内反射显微镜直接可视化单个 EV 的标记表达，并分析它们的共定位，以研究 EV 亚群。单细胞成像和共定位分析成功地说明了 EV 的多样性，并揭示了四跨膜蛋白表达的不同模式。该分析方法的应用证明了从不同常规 EV 分离方法获得的 EV 分数之间的相似性和差异性。本研究中开发的分析方法将为研究单个 EV 特性提供一种新的可靠工具，并且分析结果可能会增加对 EV 特性的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/44c36dab7087/JEV2-10-e12047-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/6b78a9ab50b5/JEV2-10-e12047-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/bcaf2ab27a2d/JEV2-10-e12047-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/f80847d46a03/JEV2-10-e12047-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/faf1722a2ffd/JEV2-10-e12047-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/c789fa919ba8/JEV2-10-e12047-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/0c04a851b55a/JEV2-10-e12047-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/44c36dab7087/JEV2-10-e12047-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/6b78a9ab50b5/JEV2-10-e12047-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/bcaf2ab27a2d/JEV2-10-e12047-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/f80847d46a03/JEV2-10-e12047-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/faf1722a2ffd/JEV2-10-e12047-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/c789fa919ba8/JEV2-10-e12047-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/0c04a851b55a/JEV2-10-e12047-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d70/7797949/44c36dab7087/JEV2-10-e12047-g007.jpg

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