E NDE fraction was smaller than the pool of all exosomes combined. Further, SEVs from all depressed patients had been considerably smaller sized than controls irrespective from the fractions. Our sequencing final results showed anOWP3.02=PT09.Immunocapturing of tumour-derived extracellular vesicles on micropatterned and antibody-conjugated surfaces for person correlative light, probe and electron measurements Pepijn Beekmana, Agustin Enciso-Martinezb, Cees Ottob and S erine Le Gacc Wageningen University, Wageningen, Netherlands; bMedical Cell Biophysics, University of Twente, Enschede, Netherlands; cApplied Microfluidics for BioEngineering Investigation, University of Twente, The Netherlands, Enschede, NetherlandsaIntroduction: Tumor-derived extracellular vesicules (tdEVs) are promising biomarkers for cancer patient management. The screening of blood samples for tdEVs shows prognostic power comparable to screening of tumour cells. However, as a result of the overlap in size RORα custom synthesis amongst tdEVs, non-cancer EVs, lipoproteins and cell debris, new approaches, not simply determined by size, are required for the reliable isolation of tdEVs and their quantification. We report an integrated evaluation methodology to study single tdEVs applying correlative data from scanning electron microscopy (SEM), Raman imaging and atomic force microscopy (AFM) to get a comprehensive dataset enabling identifying capabilities special to tdEVs. Approaches: Indium tin oxide (ITO)-coated fused silica was chosen for its low Raman background. Substrates (1 1 cm2) featuring position-dependent markings (“navigation marks”) patterned by photolithography had been modified having a monolayer of amino dodecyl phosphonic acid. The amine moieties were next reacted with poly(ethylene glycol) diglycidyl ether, forming an anti-biofouling layer. Anti-EpCAM antibodies were subsequently covalently bound on this surface. Samples of each tdEVs obtained from LNCaP cell lines and RBC-derived EVs were then introduced toJOURNAL OF EXTRACELLULAR VESICLESthe surfaces. Finally, non-specifically bound EVs had been washed away just before SEM, AFM and Raman measurements had been performed. Outcomes: Various objects were captured on the completely functionalized ITO surfaces, based on SEM imaging, although in adverse manage experiments (lacking functionalization or lacking antibody or making use of EpCAM-negative EVs), no object was detected. Principal element evaluation 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) within the captured tdEVs. AFM showed a surface coverage of 4 105 EVs per mm2 having a size PAK2 list distribution similar to that found by NTA. Summary/Conclusion: A platform was created for multi-modal analysis of selectively isolated tdEVs for their multimodal evaluation. In the future, the scope of this platform will probably be extended to other combinations of probe, light and electron microscopy approaches to relate extra parameters describing the captured EVs. Funding: Funded by NWO Perspectief.OWP3.03=PT09.The improvement 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 Well being, Bethesda, USA; b Translational Nanobiology Lab, Laboratory of Pathology, National Cancer Institute, National Institutes of Overall health, Bethesda, USAaequipped to manage large information sets compris.