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첨단연성물질 연구단
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Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis

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Title
Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis
Author(s)
Kim, Chi-Ju; Kuczler, Morgan D.; Dong, Liang; Junyoung Kim; Amend, Sarah R.; Yoon-Kyoung Cho; Pienta, Kenneth J.
Publication Date
2022-02
Journal
Journal of Visualized Experiments, v.2022, no.180
Publisher
MYJoVE Corporation
Abstract
© 2022 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.There is a need for practical assays to visualize and quantify the cells' extracellular vesicle (EV) uptake. EV uptake plays a role in intercellular communication in various research fields; cancer biology, neuroscience, and drug delivery. Many EV uptake assays have been reported in the literature; however, there is a lack of practical, detailed experimental methodology. EV uptake can be assessed by fluorescently labeling EVs to detect their location within cells. Distinguishing between internalized EVs in cells and the superficial EVs on cells is difficult, yet critical, to accurately determine the EV uptake. Therefore, an assay that efficiently quantifies EV uptake through three-dimensional (3D) fluorescence confocal microscopy is proposed in this work. Fluorescently labeled EVs were prepared using a nano-filtration-based microfluidic device, visualized by 3D confocal microscopy, and then analyzed through advanced image-processing software. The protocol provides a robust methodology for analyzing EVs on a cellular level and a practical approach for efficient analysis.
URI
https://pr.ibs.re.kr/handle/8788114/11366
DOI
10.3791/62836
ISSN
1940-087X
Appears in Collections:
Center for Soft and Living Matter(첨단연성물질 연구단) > 1. Journal Papers (저널논문)
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