Viding new insights. One example is, non-specific labelling of antibodies to lipoproteins collectively with variations in lipoprotein concentrations emphasize the relevance of fasting ahead of venipuncture. Our upcoming step is always to lengthen the software with machine understanding. Funding: NWO-TTW VENIJOURNAL OF EXTRACELLULAR VESICLESPS08.10=OWP2.Standard, high-resolution and imaging movement cytometry: potentials, pitfalls and remedies for EV characterization Jaco Botha, Rikke Wehner Rasmussen, Mathilde Sanden and Aase Handberg Division of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkpresentation give useful techniques 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 favored system for characterizing EVs, however their modest dimension have constrained the applicability of typical FCM to some extent. As a result, high-resolution and imaging FCMs are actually created but not however systematically evaluated. The aim of this presentation should be to describe the applicability of high-resolution and imaging FCM in the context of EV characterization and also the most major pitfalls potentially influencing information interpretation. Techniques: 1st, we present a side-by-side comparison of 3 unique cytometry platforms on characterizing EVs from blood plasma with regards to sensitivity, resolution and reproducibility: a conventional FCM, a high-resolution FCM and an imaging FCM. Following, we demonstrate how distinctive pitfalls can CD66e/CEACAM5 Proteins Purity & Documentation influence the interpretation of results on the different cytometry platforms. Lastly, we propose controls, answers or workarounds for knowing and limiting the influence of each of these pitfalls. Outcomes: (one) High-resolution FCM and imaging FCM displayed higher sensitivity and resolution compared to conventional FCM when measuring a mixture of nanospheres. Equally, the two solutions could detect bigger concentrations of distinct EV phenotypes than standard FCM, wherever imaging FCM outperformed highresolution FCM. Inside of day variability (n = twenty aliquots) was very similar for typical and high-resolution FCM, even though imaging FCM had a markedly greater variability. Concerning day variability (n = 5 five aliquots) was related for all three platforms. (two) The 3 most considerable pitfalls variably influencing interpretation of outcomes to the three platforms are non-specific binding of labels, antibody aggregates and entities within the sample (i.e. lipoproteins) binding EV-defining dyes. (three) By far the most important tactics for circumventing these pitfalls are stringent matching, gating and comparison of antibodies and isotype controls, high-speed centrifugation of antibodies and labels just before CD152/CTLA-4 Proteins manufacturer staining, and the use and interpretation of stained buffer controls and detergent-treated samples. Summary/conclusion: High-resolution and imaging FCM hold fantastic probable for EV characterization. Having said that, enhanced sensitivity also prospects to new artefacts and pitfalls. The solutions proposed in thisUniversity of Twente, Enschede, Netherlands; bMedical Cell Biophysics, University of Twente, Enschede, NetherlandsIntroduction: Raman spectroscopy probes molecular vibration and therefore reveals chemical data of a sample without having labelling. This optical technique might be made use of to study the chemical composition of varied EVs subtypes. EVs possess a complicated chem.