Ng. Procedures: A FACSCanto (Becton Dickinson) was adapted by replacing the 20 mW laser with a 20200 mW adjustable energy laser (each 488 nm Sapphire, Coherent). Confocal detection was achieved by replacing the common 1000 pinhole on SSC by a 200 pinhole, along with the normal photodiode on FSC by a 350 pinhole and PMT. The improvements in scatter sensitivity were quantified by calculating the scatter stain index (SI) (median intensity of a bead minus median intensity from the noise divided by two times the standard deviation from the noise) of a 500 nm polystyrene bead and the robust coefficient of variation (rCV) of a one hundred nm polystyrene bead (both BioCytex). Ideally the SI is as higher as you can and rCV as low as you can.JOURNAL OF EXTRACELLULAR VESICLESResults: A 10-fold increase in laser power Parathyroid Hormone Receptor Proteins MedChemExpress elevated the SI on SSC 2.9-fold and on FSC 20-fold, whereas the rCV enhanced (lowered 0.67-fold and 0.97-fold, respectively). The enhanced confocal detection elevated the SI on SSC 6.4-fold and on FSC 550fold, while the rCV slightly worsened (improved 1.1fold and 1.02-fold, respectively). Combining both increased laser power and confocal detection resulted inside a 20-fold increase in SI for SSC and 2 10^4-fold for FSC, and improved the rCV (lowered 0.39-fold and 0.24-fold, respectively). Summary/Conclusion: Adaption from the optical configuration with the FACSCanto by escalating the laser power and confocal detection enhanced the scatter sensitivity 20-fold for SSC and 2 10^4-fold for FSC. Subsequent, we will evaluate the influence of improved measurement time and reduction of the number of particles in the sheath on the scatter sensitivity. Funding: NWO-TTW Perspectief CANCER-IDPF06.Lipoprotein particles could be detected by high-resolution flow cytometry and potentially interfere with EV characterisation Rikke Wehner Rasmussen, Jaco Botha, Mathilde Sanden and Aase Handberg Division of Clinical Biochemistry, CD1b Proteins Recombinant Proteins Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkphenotypes. From 0 FT cycles, ApoB bound to PS +CD41+ and PS+CD41+ CD36+ phenotypes tended to decrease (p 0.05). Additionally, ApoB bound to PS +CD36+ improved four.9-fold from 0 FT cycles for (p 0.05). Interestingly, this progression mirrored that of PS+CD36+ (two.0.5-fold, p 0.05), bulk CD36 + (1.8.4-fold, p 0.05) and ApoB+ (4.1.0-fold, p 0.01). Ultimately, in line with earlier reports, PS+ tended to enhance following FT (1.5-2.1-fold, p 0.05). Contrary to previous reports, certain EV phenotypes decreased from 0 FT cycles (PS+CD41+ and PS +CD41+ CD36+, each two.6-fold, p 0.05) suggesting that EV phenotypes may perish following FT additional confirmed on bi-variable plots of information. Summary/Conclusion: This study demonstrates that ApoB is usually detected on hFCM and thereby interfere with EV characterisation. What additional complicates matters is that lipoproteins could carry markers traditionally connected with EVs which includes PS and CD36. FT cycles didn’t regularly dissociate EVs and lipoproteins; however, FT affected certain EV populations. Additional studies are required to elucidate these findings.PF06.Analysis of fluorescent labelling efficiency of extracellular vesicles derived from diverse kingdoms of life with lipid-binding dyes through nano-flow cytometry Ye Tiana, Chen Chenb, Qian Niua, Shaobin Zhuc and Xiaomei Yand Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, China, Xiamen, China (People’s Republic); bInstitute for Chemical Resear.