E NDE fraction was smaller than the pool of all exosomes LFA-3/CD58 Proteins Source combined. Additional, SEVs from all depressed individuals had been considerably smaller sized than controls irrespective in the fractions. Our sequencing benefits showed anOWP3.02=PT09.Immunocapturing of tumour-derived extracellular vesicles on micropatterned and SR-BI/CD36 Proteins custom synthesis 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 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 power comparable to screening of tumour cells. On the other hand, on account of the overlap in size in between tdEVs, non-cancer EVs, lipoproteins and cell debris, new approaches, not merely determined by size, are necessary for the trustworthy isolation of tdEVs and their quantification. We report an integrated analysis methodology to study single tdEVs utilizing correlative information from scanning electron microscopy (SEM), Raman imaging and atomic force microscopy (AFM) to receive a comprehensive dataset enabling identifying features exceptional to tdEVs. Techniques: 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 have been modified having a monolayer of amino dodecyl phosphonic acid. The amine moieties had been next 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 each tdEVs obtained from LNCaP cell lines and RBC-derived EVs were then introduced toJOURNAL OF EXTRACELLULAR VESICLESthe surfaces. Ultimately, non-specifically bound EVs have been washed away before SEM, AFM and Raman measurements were performed. Outcomes: A number of objects had been captured on the fully functionalized ITO surfaces, based on SEM imaging, when in unfavorable handle experiments (lacking functionalization or lacking antibody or utilizing 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) in the captured tdEVs. AFM showed a surface coverage of four 105 EVs per mm2 using a size distribution related to that located by NTA. Summary/Conclusion: A platform was created for multi-modal evaluation of selectively isolated tdEVs for their multimodal analysis. Within the future, the scope of this platform will be extended to other combinations of probe, light and electron microscopy tactics 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 Overall health, Bethesda, USA; b Translational Nanobiology Lab, Laboratory of Pathology, National Cancer Institute, National Institutes of Overall health, Bethesda, USAaequipped to deal with large information sets compris.
