Fig. 5.

SepMate density gradient centrifugation yields less PBMCs but does not influence cell viability or proportion of T and B cells compared with Ficoll-Paque. a–f Peripheral blood was collected from six healthy donors, and equal volumes were used to separate PBMCs by Ficoll-Paque and SepMate density gradient centrifugation. c Cell numbers and d cell viabilities were assessed using trypan blue and a haemocytometer. a, b, e, f Cells were labelled with fluorochrome-conjugated mAbs and data acquired by flow cytometry. a, b Sequential flow cytometric gates were selected left to right, as shown, to gate single cells, lymphocytes, live (Zombie NIR⁻) CD45⁺ lymphocytes, in order to determine proportions of e CD3⁺CD19⁻ T cells and f CD3⁻CD19⁺ B cells amongst CD45⁺ cells. a, b A representative gating strategy of PBMCs isolated by a Ficoll-Paque or b SepMate density gradient centrifugation is shown. c, e, f Connected or d individual symbols represent individual donors (n = 6); P < 0.05 indicates significance; ns, no significances; paired two-tailed Student’s t test