Detecting individual extracellular vesicles using a multicolor in situ proximity ligation assay with flow cytometric readout
Autor: | K. Göran Ronquist, Tonge Ebai, Ulf Landegren, Lotta Wik, Louise Dubois, Olivia Nolander, Masood Kamali-Moghaddam, Liza Löf, Ola Söderberg, Emma Lundin |
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Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
In situ Multidisciplinary medicine.diagnostic_test Cell- och molekylärbiologi Proximity ligation assay Biology Extracellular vesicles Molecular medicine Article Flow cytometry Cell biology 03 medical and health sciences Cell and molecular biology 030104 developmental biology 0302 clinical medicine Qualitative analysis Flow (mathematics) 030220 oncology & carcinogenesis medicine Cell and Molecular Biology |
Zdroj: | Scientific Reports |
ISSN: | 2045-2322 |
Popis: | 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. |
Databáze: | OpenAIRE |
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