Ng. Techniques: A FACSCanto (Becton Dickinson) was adapted by replacing the 20 mW laser with a 20200 mW adjustable energy laser (both 488 nm Sapphire, Coherent). Confocal detection was achieved by replacing the normal 1000 pinhole on SSC by a 200 pinhole, and also the typical 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 of the noise divided by two times the regular deviation with the noise) of a 500 nm polystyrene bead plus the robust coefficient of variation (rCV) of a one hundred nm polystyrene bead (each 5-HT6 Receptor Modulator drug BioCytex). Ideally the SI is as higher as you can and rCV as low as possible.JOURNAL OF EXTRACELLULAR VESICLESResults: A 10-fold increase in laser power increased the SI on SSC two.9-fold and on FSC 20-fold, whereas the rCV enhanced (decreased 0.67-fold and 0.97-fold, respectively). The improved confocal detection increased the SI on SSC 6.4-fold and on FSC 550fold, although the rCV slightly worsened (enhanced 1.1fold and 1.02-fold, respectively). Combining both elevated laser power and confocal detection resulted within a 20-fold improve in SI for SSC and two 10^4-fold for FSC, and enhanced the rCV (reduced 0.39-fold and 0.24-fold, respectively). Summary/Conclusion: Adaption in the optical configuration on the FACSCanto by increasing the laser energy and confocal detection improved the scatter sensitivity 20-fold for SSC and two 10^4-fold for FSC. Subsequent, we’ll evaluate the influence of enhanced measurement time and reduction from the PLK1 Molecular Weight number of particles within the sheath on the scatter sensitivity. Funding: NWO-TTW Perspectief CANCER-IDPF06.Lipoprotein particles might be detected by high-resolution flow cytometry and potentially interfere with EV characterisation Rikke Wehner Rasmussen, Jaco Botha, Mathilde Sanden and Aase Handberg Department of Clinical Biochemistry, Aalborg University Hospital, Aalborg, Denmark, Aalborg, Denmarkphenotypes. From 0 FT cycles, ApoB bound to PS +CD41+ and PS+CD41+ CD36+ phenotypes tended to reduce (p 0.05). In addition, ApoB bound to PS +CD36+ elevated four.9-fold from 0 FT cycles for (p 0.05). Interestingly, this progression mirrored that of PS+CD36+ (2.0.5-fold, p 0.05), bulk CD36 + (1.8.4-fold, p 0.05) and ApoB+ (4.1.0-fold, p 0.01). Lastly, in line with earlier reports, PS+ tended to increase following FT (1.5-2.1-fold, p 0.05). Contrary to previous reports, particular EV phenotypes decreased from 0 FT cycles (PS+CD41+ and PS +CD41+ CD36+, each 2.6-fold, p 0.05) suggesting that EV phenotypes may well perish following FT additional confirmed on bi-variable plots of information. Summary/Conclusion: This study demonstrates that ApoB is often detected on hFCM and thereby interfere with EV characterisation. What further complicates matters is that lipoproteins could carry markers traditionally associated with EVs including PS and CD36. FT cycles did not regularly dissociate EVs and lipoproteins; however, FT affected particular EV populations. Further studies are needed to elucidate these findings.PF06.Analysis of fluorescent labelling efficiency of extracellular vesicles derived from various kingdoms of life with lipid-binding dyes via nano-flow cytometry Ye Tiana, Chen Chenb, Qian Niua, Shaobin Zhuc and Xiaomei Yand Division of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, China, Xiamen, China (People’s Republic); bInstitute for Chemical Resear.
