A Simplified Workflow for Flow Cytometric Analysis of Extracellular Vesicles on the Invitrogen™ Attune™ NxT Flow Cytometer

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Presentation Time: 11:30 AM - 12:45 PM
Poster Board Number: B883
Abstract ID: 4556

Presenting Author:
Samantha Ellis , Scientist III at Thermo Fisher Scientific

Abstract:

Extracellular vesicles (EV) are miniscule particles composed of a bilipid membrane that are released by all cells. In recent years, it has become clear that EV play an important role in cell-cell communication by shuttling their cargo of proteins, lipids, and nucleic acids. As their content can be indicative of the health of their cell of origin and ability to deliver functional biomolecules, EV have become an increasingly popular topic of research for their diagnostic and therapeutic potential.

EV vary in size, surface markers, and content. Their heterogenous nature introduces challenges for analysis when using technologies that were developed for cells. In flow cytometry, EV have a lower refractive index and paucity of epitopes comparative to cells, which hampers detection. In addition, few commercially available antibodies have been specifically validated for EV flow. This combined with the need for tedious isolation processes and a limited range of instruments capable of small particle detection can make EV flow cytometry a daunting endeavor.

Despite this, flow cytometry holds promise as an accessible approach to analyze EV. Here, we present a streamlined protocol of EV characterization by flow cytometry in which we adapted the Invitrogen™ CELLection™ Pan Mouse IgG Kit for EV capture and staining and optimized the Invitrogen™ Attune™ NxT Flow Cytometer for small particle resolution to validate monoclonal antibodies targeting EV from human plasma. For research use only.