Rrelative information from scanning electron microscopy (SEM), Raman imaging (RI) and atomic force microscopy (AFM) to obtain a complete dataset permitting identifying attributes exclusive to tdEVs. Solutions: Indium tin oxide (ITO)-coated fused silica was selected for its low Raman background. Substrates (1 x 1 cm2) featuring position-dependent markings (“navigation marks”) patterned by photolithography had been modified with a monolayer of amino dodecyl phosphonic acid. The amine moieties had been subsequent reacted with poly(ethylene glycol) diglycidyl ether, forming an anti-biofouling layer. Anti-EpCAM antibodies have been subsequently covalently bound on this surface. Samples of both tdEVs obtained from LNCaP cell lines and RBC-derived EVs had been then introduced to the surfaces. Lastly, non-specifically bound EVs had been washed away just before SEM, AFM and Raman measurements had been performed. Results: Numerous objects had been captured on the totally functionalized ITO surfaces, as outlined by SEM imaging, though in negative handle experiments (lacking functionalization or lacking antibody or applying EpCAM-negative EVs), no object was detected. Principal element analysis of their Raman spectra, previously demonstrated to be in a position to distinguish tdEVs from RBC-derived EVs, revealed the presence of characteristic lipid bands (e.g. 2851 cm-1) in the captured tdEVs. AFM showed a surface coverage of ,four 10^5 EVs per mm2 using a size distribution related to that identified by NTA. Summary/conclusion: A platform was developed for multi-modal analysis of selectively isolated tdEVs for their multi-modal evaluation. In the future, the scope of this platform will probably be extended to other combinations of probe, light and electron microscopy tactics to relate further parameters describing the captured EVs. Funding: Funded by NWO PerspectiefWageningen University, Wageningen, Netherlands; bMedical Cell Biophysics, University of Twente, CD25/IL-2R alpha Proteins Source Enschede, Netherlands; cApplied Microfluidics for BioEngineering Research, University of Twente, The Netherlands, Enschede, NetherlandsPT09.14=OWP3.The development of a scalable extracellular vesicle subset characterization pipeline. Joshua Welsha, Julia Kepleyb and Jennifer C. Jonesa Translational Nanobiology Section, Laboratory of Pathology, National Cancer Institute, National Institutes of Overall health, Bethesda, USA; b Translational Nanobiology Lab, Laboratory of Pathology, National Cancer Institute, National Institutes of Overall health, Bethesda, USAaIntroduction: Tumour-derived extracellular vesicles (tdEVs) are promising biomarkers for cancer patient management. The screening of blood samples for tdEVs shows prognostic power comparable to screening of tumour cells. Even so, resulting from the overlap in size in between tdEVs, non-cancer EVs, lipoproteins and cell debris, new approaches, not merely according to size, are essential for the reliable isolation of tdEVs and their quantification. We report an integrated analysisIntroduction: Liquid biopsies offer you an important alternative to tumour biopsies that might be restricted by the challenges of invasive procedures. We hypothesize thatISEV2019 ABSTRACT BOOKcirculating Extracellular Vesicles (EVs) and their cargo might present a BTN1A1 Proteins custom synthesis helpful surrogate biopsy strategy. Resulting from their modest diameter (30-1000 nm), EVs migrate from tissue in to the peripheral circulation and offer a snapshot in the generating cells. Our lab has created a first-in-class pipeline to work with single cell omics techniques to characterize EV heterogeneity with high-sensitivity by combining mu.
