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. 2015 Oct 1;87:64–74. doi: 10.1016/j.ymeth.2015.03.028

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© 2015 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Fig. 3 — Representative flow cytometric analyses of fractionated maternal side placental perfusate. (Ai) Forward scatter (FSC) vs. side scatter (SSC) profile of maternal side placental perfusate, (Aii) histogram showing 100% labelling of red blood cells (RBC), (Aiii) maternal side placental perfusate supernatant (1500SN) following depletion of contaminating RBC by 2 × 1500×g centrifugation for 10 min, (Aiv) 1500SN bio-maleimide positive labelling, (Av) FSC vs. SSC profile of 10,000×g pellet (10KP) and (Avi) 10KP bio-maleimide positive labelling. (Bi) 1500SN bio-maleimide positive EV displayed on two-parameter plots and phenotype identified: syncytiotrophoblast derived EV (STBEV), PLAP⁺/HLA Class I⁻, (Bii) RBC EV, CD235a/b⁺/PLAP⁻, (Biii) platelet EV (CD41⁺/PLAP⁻), (Biv) leukocyte/endothelial EV (HLA Class I⁺/CD41⁻). (Ci) 10KP bio-maleimide positive EV displayed on two-parameter plots and phenotype identified: STBEV, PLAP⁺/HLA Class I⁻, (Cii) RBC EV, CD235a/b⁺/PLAP⁻, (Ciii) platelet EV (CD41⁺/PLAP), (Civ) leukocyte/endothelial EV (HLA Class I⁺/CD41⁻). Histograms show the mean ± SE for percentage positive EV and two-parameter plots show the mean for percentage positive EV (n = 8). Percentages have been adjusted to account for background contaminating particulates.