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Detecting individual extracellular vesicles using a multicolor in situ proximity ligation assay with flow cytometric readout
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Biochemial structure and function.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Molecular tools. Uppsala University, Science for Life Laboratory, SciLifeLab.
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2016 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 6, 34358Article in journal (Refereed) Published
Abstract [en]

Flow cytometry is a powerful method for quantitative and qualitative analysis of individual cells. However, flow cytometric analysis of extracellular vesicles (EVs), and the proteins present on their surfaces has been hampered by the small size of the EVs - in particular for the smallest EVs, which can be as little as 40 nm in diameter, the limited number of antigens present, and their low refractive index. We addressed these limitations for detection and characterization of EV by flow cytometry through the use of multiplex and multicolor in situ proximity ligation assays (in situ PLA), allowing each detected EV to be easily recorded over background noise using a conventional flow cytometer. By targeting sets of proteins on the surface that are specific for distinct classes of EVs, the method allows for selective recognition of populations of EVs in samples containing more than one type of EVs. The method presented herein opens up for analyses of EVs using flow cytometry for their characterization and quantification.

Place, publisher, year, edition, pages
2016. Vol. 6, 34358
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-306259DOI: 10.1038/srep34358ISI: 000384185900001PubMedID: 27681459OAI: oai:DiVA.org:uu-306259DiVA: diva2:1040214
Funder
EU, European Research Council, 259796; 294409Swedish Research Council
Available from: 2016-10-26 Created: 2016-10-26 Last updated: 2016-10-26Bibliographically approved

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Löf, LizaEbai, TongeDubois, LouiseRonquist, K. GöranSöderberg, OlaLandegren, UlfKamali-Moghaddam, Masood
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