Deep phenotyping of peripheral mononuclear blood cells in health and disease
Peripheral blood mononuclear cells (PBMCs) were isolated from healthy controls and patients with various diseases.. Cells were stained with CD45 and sample tags for identification purposes. CD45+ live cells were enriched through flow sorting and samples were pooled. Samples were then stained with 30 AbSeq antibodies and loaded onto 4 cartridges to account for interplate differences. Pooled sequencing libraries were then sequenced on NovaSeq 6000 (Illumina) using a S2 Reagent Kit v1.5 (200 cycles). Overall design: For human scRNAseq, PBMCS were isolated from healthy controls and patients with various diseases. Cells were stained with CD45 and sample tags. CD45+ live cells were enriched through flow sorting and samples were pooled. Samples were then stained with 30 AbSeq antibodies and loaded onto 4 cartridges and subjected to Rhapsody analysis following manufacturers protocol. Pooled sequencing libraries were then sequenced on NovaSeq 6000 (Illumina) using a S2 Reagent Kit v1.5 (200 cycles).