Viding new insights. As an example, non-specific labelling of antibodies to lipoproteins collectively with variations in lipoprotein concentrations emphasize the relevance of fasting ahead of venipuncture. Our upcoming stage is to extend the software program with machine discovering. Funding: NWO-TTW VENIJOURNAL OF EXTRACELLULAR VESICLESPS08.10=OWP2.Traditional, high-resolution and imaging movement cytometry: potentials, pitfalls and solutions for EV characterization Jaco Botha, Rikke Wehner Rasmussen, Mathilde Sanden and Aase Handberg Division of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkpresentation present practical 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: Movement cytometry (FCM) has prolonged been a favored technique for characterizing EVs, having said that their little size have constrained the applicability of standard FCM to some extent. Consequently, high-resolution and imaging FCMs have already been produced but not yet systematically evaluated. The aim of this presentation is usually to describe the applicability of high-resolution and imaging FCM from the context of EV characterization plus the most significant pitfalls probably influencing data interpretation. Procedures: Initial, we present a side-by-side comparison of three various cytometry platforms on characterizing EVs from blood plasma with regards to sensitivity, resolution and reproducibility: a Oxytocin Proteins Synonyms typical FCM, a high-resolution FCM and an imaging FCM. Subsequent, we demonstrate how various pitfalls can influence the interpretation of effects about the various cytometry platforms. Lastly, we propose controls, remedies or workarounds for understanding and limiting the influence of each of these pitfalls. Outcomes: (one) High-resolution FCM and imaging FCM displayed greater sensitivity and resolution compared to conventional FCM when measuring a mixture of nanospheres. Equally, both approaches could detect greater concentrations of unique EV phenotypes than standard FCM, the place imaging FCM outperformed highresolution FCM. Inside of day variability (n = twenty aliquots) was similar for conventional and high-resolution FCM, though imaging FCM had a markedly bigger variability. Involving day variability (n = five 5 aliquots) was very similar for all 3 platforms. (2) The 3 most substantial pitfalls variably influencing interpretation of outcomes around the 3 platforms are non-specific binding of labels, antibody aggregates and entities in the sample (i.e. lipoproteins) binding EV-defining dyes. (three) One of the most important methods for circumventing these pitfalls are stringent matching, gating and comparison of antibodies and isotype controls, high-speed centrifugation of antibodies and labels before staining, and the use and interpretation of stained buffer controls and detergent-treated samples. Summary/conclusion: High-resolution and imaging FCM hold excellent potential for EV characterization. Even so, greater sensitivity also leads to new artefacts and pitfalls. The remedies proposed in thisUniversity of Twente, CD45 Proteins medchemexpress Enschede, Netherlands; bMedical Cell Biophysics, University of Twente, Enschede, NetherlandsIntroduction: Raman spectroscopy probes molecular vibration and thus reveals chemical information and facts of a sample without labelling. This optical approach can be employed to research the chemical composition of various EVs subtypes. EVs possess a complicated chem.