Vital for precise and complete characterization of EVs in biological samples with good reproducibility. References 1. Obeid et al., B B. 2017. 93:25059 2. Obeid et al., NBM. 2019 (in revision) 3. Thery et al., JEV. 2018. 8;1535750 Funding: Region Franche-Comt2017020.PT09.Multi-parametric single vesicle evaluation applying an interferometric imaging platform George Daaboula, Veronica Sanchezb, Aditya Dhandeb, Chetan Soodb, Gregg Lithgowb, Francis Fordjourc, Stephen Gouldc and David Freedmanba NanoView Biosciences, Brighton, USA; bNanoView Biosciences, Boston, USA; cJohn Hopkins University, Baltimore, USAThe calculated fluorescence detection limit approaches single fluorescence sensitivity established using fluorescent polystyrene nanoparticles (2000nm diameter) corresponding to 18010,000 MESF. Final results: A tetraspanin assay was created around the ExoViewTM platform for the detection of CD81, CD63, CD9 positive vesicles directly from cell culture samples without having the require for purification. We are able to also permeabilize the vesicles to probe the cargo of person vesicles. To validate the detection of Glycophorin-A/CD235a Proteins web tetraspanins and internal cargo proteins we applied knockout cell lines as damaging controls. The assay can also be utilized for detection of vesicles from other biological fluids like urine, plasma, CSF, and saliva. We demonstrated that most tetraspanin optimistic vesicles have a diameter around 50 nm, which agrees with TEM, versus the extensively reported diameter of 100nm within the literature. Summary/Conclusion: The ExoView platform is a scalable single vesicle analysis platform which can size, enumerate and run multi-parametric co-localization experiments straight from sample. The platform could be applied for simple investigation also as biomarker discovery for liquid-biopsy applications.PT09.Quantification of circulating extracellular vesicles from human plasma by using a membrane-based microfluidic method Yi-Sin Chena, Gwo-Bin Leea and Chihchen ChenbaIntroduction: Existing single vesicle analysis tactics like electron microscopy and atomic force microscopy require high expertise and are restricted in throughput. Flow cytometry (FC), which is on a regular basis employed to for single cell evaluation and sorting, has restricted sensitivity in light scatter mode for detection of very abundant populations of EVs smaller sized than a 100 nm. Recent publications show that the exosome typical diameter is about 50 nm, which has been measured by super-resolution imaging, nanoFCM, and TEM. The additional sensitive fluorescence-based detection of EVs is also difficult mainly because EVs could have substantially much less than ten epitopes of your marker of interest, a limit for most FC systems. Solutions: To address the limitation in single vesicles analysis we’ve developed a method that will size, enumerate, and co-localize 4 markers (surface and cargo) on single vesicles across 10 distinct Trk receptors Proteins site subpopulations on a single sensor surface. The approach is termed SP-IRIS and commercialized as ExoViewTM by NanoView Biosciences. EvoViewTM relies on a bilayer substrate (silicon/silicon dioxide) that forms a typical path interferometer for enhanced nanoparticle evaluation.Department of Power Mechanical Engineering, National Tsing Hua University, Taiwan, Hsinchu, Taiwan (Republic of China); bInstitution of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan (Republic of China)Introduction: Extracellular vesicles (EVs) have served as biomarkers for cancer diagnosis and prognosis based on their carried.