Often regarded as a вЂњsnapshotвЂќ of the status of the cell of origin, EVs are examined for their biochemical signatures to assess the presence of various diseases, e.g., cancer or viral infections 7, 8, and are considered excellent minimally invasive biomarkers in so-called liquid biopsies 9, 10, 11. As EVs are excreted by virtually all cell types in the human body, they can be found in most body fluids, such as the blood 1, saliva 4 and urine 5, 6. They are involved in cellular communication through transfer of surface receptors and/or a variety of macromolecules carried as cargo (e.g., lipids, proteins, nucleic acids, protein-coding mRNAs and regulatory microRNAs) 2, 3. Additionally, this direct approach for EV analysis will enable researchers to explore corners of EVs as cellular messengers in healthy and pathological conditions.Д®xtracellular vesicles (EVs) are lipid bilayer membrane structures (30вЂ“8000вЂ‰nm in diameter 1) released by cells. Through this work, we provide a framework that will allow the application of IFCM for EV analysis in peripheral blood plasma in a plethora of experimental and potentially diagnostic settings. Using a combination of markers (CFSE & Tetraspanins, or CD9 & CD31) we found that >90% of double-positive fluorescent events represented single EVs. Detergent treatment of EVs was performed to discriminate true vesicular events from artefacts. Instrument calibration (both size and fluorescence) were performed with commercial polystyrene beads.

To rule out selection bias or introduction of artefacts caused by EV isolation techniques, we present a clinically feasible, imaging flow cytometry (IFCM)вЂ“based methodology to phenotype and determine the concentration of EVs with a diameter в‰¤400вЂ‰nm in human platelet-poor plasma (PPP) without prior isolation of EVs.

Extracellular vesicles (EVs) are tissue-specific particles released by cells containing valuable diagnostic information in the form of various biomolecules.

Core shell column