Viding new insights. By way of example, 5-HT7 Receptor Modulator supplier non-specific labelling of antibodies to lipoproteins together with variations in lipoprotein concentrations emphasize the relevance of fasting just before venipuncture. Our upcoming step should be to extend the software program with machine finding out. Funding: NWO-TTW VENIJOURNAL OF EXTRACELLULAR VESICLESPS08.10=OWP2.Typical, high-resolution and imaging movement cytometry: potentials, pitfalls and solutions for EV characterization Jaco Botha, Rikke Wehner Rasmussen, Mathilde TLR3 Source Sanden and Aase Handberg Department of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkpresentation offer handy strategies for circumventing these.PS08.11=OWP2.Convolutional neural networks for classification of tumour derived extracellular vesicles Wooje Leea, Aufried Lenferinka, Cees Ottob and Herman OfferhausaaIntroduction: Flow cytometry (FCM) has prolonged been a preferred method for characterizing EVs, having said that their modest dimension have constrained the applicability of traditional FCM to some extent. Consequently, high-resolution and imaging FCMs have already been created but not but systematically evaluated. The aim of this presentation is to describe the applicability of high-resolution and imaging FCM within the context of EV characterization and also the most important pitfalls probably influencing information interpretation. Approaches: First, we current a side-by-side comparison of three distinctive cytometry platforms on characterizing EVs from blood plasma relating to sensitivity, resolution and reproducibility: a traditional FCM, a high-resolution FCM and an imaging FCM. Next, we show how diverse pitfalls can influence the interpretation of success on the diverse cytometry platforms. Last but not least, we propose controls, answers or workarounds for understanding and limiting the influence of every of these pitfalls. Outcomes: (one) High-resolution FCM and imaging FCM displayed higher sensitivity and resolution in contrast to standard FCM when measuring a mixture of nanospheres. Equally, each methods could detect greater concentrations of precise EV phenotypes than traditional FCM, wherever imaging FCM outperformed highresolution FCM. Inside of day variability (n = 20 aliquots) was related for conventional and high-resolution FCM, while imaging FCM had a markedly larger variability. In between day variability (n = five five aliquots) was comparable for all three platforms. (2) The 3 most significant pitfalls variably influencing interpretation of results around the 3 platforms are non-specific binding of labels, antibody aggregates and entities within the sample (i.e. lipoproteins) binding EV-defining dyes. (3) Essentially the most essential tactics for circumventing these pitfalls are stringent matching, gating and comparison of antibodies and isotype controls, high-speed centrifugation of antibodies and labels prior to staining, and the use and interpretation of stained buffer controls and detergent-treated samples. Summary/conclusion: High-resolution and imaging FCM hold terrific potential for EV characterization. Even so, enhanced sensitivity also prospects to new artefacts and pitfalls. The options proposed in thisUniversity of Twente, Enschede, Netherlands; bMedical Cell Biophysics, University of Twente, Enschede, NetherlandsIntroduction: Raman spectroscopy probes molecular vibration and as a result reveals chemical data of the sample without labelling. This optical technique is usually employed to examine the chemical composition of various EVs subtypes. EVs have a complicated chem.