E NDE fraction was smaller than the pool of all exosomes combined. Further, SEVs from all depressed individuals had been substantially smaller sized than controls irrespective of your fractions. Our sequencing outcomes showed anOWP3.02=PT09.Immunocapturing of tumour-derived extracellular vesicles on micropatterned and antibody-conjugated surfaces for individual 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 Analysis, 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 energy comparable to screening of tumour cells. Even so, because of the overlap in size among tdEVs, non-cancer EVs, lipoproteins and cell debris, new approaches, not only based on size, are essential for the reputable isolation of tdEVs and their quantification. We report an integrated evaluation methodology to study single tdEVs utilizing correlative data from scanning electron microscopy (SEM), Raman imaging and atomic force microscopy (AFM) to obtain a comprehensive dataset allowing identifying options exceptional to tdEVs. Solutions: Indium tin oxide (ITO)-coated fused silica was selected for its low Raman background. Substrates (1 1 cm2) featuring position-dependent markings (“navigation marks”) patterned by photolithography have been modified using 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 had been then introduced toJOURNAL OF EXTRACELLULAR VESICLESthe surfaces. Finally, non-specifically bound EVs had been PD-L1 Proteins Recombinant Proteins washed away prior to SEM, AFM and Raman measurements were performed. Benefits: Several objects were captured on the fully functionalized ITO surfaces, in accordance with SEM imaging, while in negative handle experiments (lacking functionalization or lacking antibody or employing EpCAM-negative EVs), no object was detected. Principal component evaluation of their Raman spectra, previously demonstrated to become able to distinguish tdEVs from RBC-derived EVs, revealed the presence of characteristic lipid bands (e.g. 2851 cm-1) Adhesion GPCRs Proteins Storage & Stability inside the captured tdEVs. AFM showed a surface coverage of 4 105 EVs per mm2 using a size distribution related to that discovered by NTA. Summary/Conclusion: A platform was developed for multi-modal analysis of selectively isolated tdEVs for their multimodal analysis. Within the future, the scope of this platform will probably be extended to other combinations of probe, light and electron microscopy approaches to relate additional 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 Wellness, Bethesda, USA; b Translational Nanobiology Lab, Laboratory of Pathology, National Cancer Institute, National Institutes of Wellness, Bethesda, USAaequipped to manage significant data sets compris.