Viding new insights. For example, non-specific labelling of antibodies to lipoproteins together with variations in

October 28, 2022

Viding new insights. For example, non-specific labelling of antibodies to lipoproteins together with variations in lipoprotein concentrations emphasize the relevance of fasting ahead of MSR1/CD204 Proteins Molecular Weight venipuncture. Our following step would be to lengthen the software program with machine understanding. Funding: NWO-TTW VENIJOURNAL OF EXTRACELLULAR VESICLESPS08.10=OWP2.Traditional, high-resolution and imaging flow cytometry: potentials, pitfalls and remedies for EV characterization Jaco Botha, Rikke Wehner Rasmussen, Mathilde Sanden and Aase Handberg Department of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkpresentation provide useful 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 approach for characterizing EVs, on the other hand their smaller size have limited the applicability of conventional FCM to some extent. So, high-resolution and imaging FCMs have already been designed but not nevertheless systematically evaluated. The aim of this presentation would be to describe the applicability of high-resolution and imaging FCM during the context of EV characterization and the most considerable pitfalls probably influencing data interpretation. Techniques: Initial, we existing a side-by-side comparison of three diverse 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. Subsequent, we show how different pitfalls can influence the CD134/OX40 Proteins Storage & Stability interpretation of benefits on the diverse cytometry platforms. Eventually, we propose controls, options or workarounds for knowing and limiting the influence of each of these pitfalls. Final results: (1) High-resolution FCM and imaging FCM displayed higher sensitivity and resolution compared to standard FCM when measuring a mixture of nanospheres. Equally, each strategies could detect greater concentrations of unique EV phenotypes than standard FCM, in which imaging FCM outperformed highresolution FCM. Inside day variability (n = twenty aliquots) was similar for typical and high-resolution FCM, even though imaging FCM had a markedly greater variability. In between day variability (n = 5 five aliquots) was very similar for all 3 platforms. (2) The three most substantial pitfalls variably influencing interpretation of results around the 3 platforms are non-specific binding of labels, antibody aggregates and entities in the sample (i.e. lipoproteins) binding EV-defining dyes. (3) One of the most important strategies 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 fantastic potential for EV characterization. Even so, increased sensitivity also prospects to new artefacts and pitfalls. The answers proposed in thisUniversity of Twente, Enschede, Netherlands; bMedical Cell Biophysics, University of Twente, Enschede, NetherlandsIntroduction: Raman spectroscopy probes molecular vibration and therefore reveals chemical information of a sample with out labelling. This optical technique could be utilised to study the chemical composition of diverse EVs subtypes. EVs possess a complicated chem.