N more option. The typical SSC detector remains in place plus the SP SSC module has minimal impact on normal SSC and fluorescent overall performance therefore use on the system for cell evaluation applications is still achievable. Initial results employing the SP SSC module had been obtained utilizing a BD FACSCelestaTM SORP as well as a BD FACSAriaTM Fusion, respectively possessing a 100 and 200 mW 488 laser. Side-by-side comparison in the regular SSC detection vs. SP SSC detection was performed making use of polystyrene beads, silica beads, EV reference material and antibodystained EV material. Summary/conclusion: Utilization with the SP SSC module for sorting of natural (plasma EVs) and artificialISEV2019 ABSTRACT BOOK(liposomes) membrane particles is presently getting undertaken.IP.IP.Benchmarking of established exosome isolation techniques (density gradient centrifugation, size-exclusion chromatography and immunebead separation) with glycan recognizing EX ead Dapi Meng Lin. Chianga, Chin-Sheng Linb and Michael Pfafflca Biovesicle; bDivision of Cardiology, Tri-Service General PIM2 Accession Hospital, Taiwan National Defense Medical Center, Taiwan; cAnimal Physiology and Immunology, College of Life Sciences Weihenstephan, Technical University of Munich, Freising, GermanyQuantitative imaging and phenotyping of EVs with 20 nm resolution Andras Miklosi, Zehra Nizami, Blanka Kellermayer and Mariya Georgieva ONI (Oxford Nanoimaging ltd)SMYD2 review Introduction: Complex extracellular vesicle (EV) phenotyping is really a significant technical challenge that hinders clinical translation. Single-molecule localization microscopy (SMLM) can be a Nobel-Prize winning method that makes it possible for quantitative imaging beneath the diffraction limit necessitating only uncomplicated and speedy sample preparation. The information presented here constitutes one of the very first accounts of single-molecule imaging made use of to effectively resolve the structure, protein (CD9, CD63, and CD81) and nucleic acid content of EVs with 20 nm resolution. Techniques: EV isolation was performed from keratinocyte culture media. EV suspensions were stained working with fluorescently labelled primary antibodies raised against recognized exosome markers, and commercially readily available membrane and nucleic acid labels. Characterization of the molecular content and structural properties of surface-immobilized EVs was performed working with the SMLM mode in the ONI Nanoimager. Sizing of EVs in remedy was performed employing the dual-colour single-particle tracking mode on the ONI Nanoimager. Final results: Multicolour super-resolution microscopy imaging of purified EVs revealed the phenotypic and structural properties of a huge selection of individual vesicles at a time. The membrane staining allowed to visualize EVs with sizes ranging from 20 nm to 250 nm, and sizing by tracking confirmed this distribution in addition to a mean size of 120 nm. For EVs of 40 nm the membrane appeared as a ring and was a confirmation of their intact structure. CD63, CD9 and CD81 co-localized with the membrane staining in the nm scale, hence permitting to decide the molecular ID of EV subpopulations and correlate the protein marker levels together with the size of EVs. Summary/conclusion: The quantitative nature of single-molecule imaging and tracking significantly improves EV characterization. This work supplies evidence in the use of SMLM imaging as a novel and highly effective tool for fast and multiplexed EV characterization with exclusive mixture of structural and phenotypic insight.Introduction: Exosomes are compact vesicles (30150 nm) discovered in a variety of human biofluids, including.