Fig. 1: Workflow to link IgG secretion to surface markers and transcriptomes at the single-cell level. | Nature Communications

Fig. 1: Workflow to link IgG secretion to surface markers and transcriptomes at the single-cell level.

From: SEC-seq: association of molecular signatures with antibody secretion in thousands of single human plasma cells

Fig. 1

Human B cells are isolated from donors and expanded in a differentiation cocktail to promote differentiation into antibody-secreting cells. Cells are then loaded into a slurry of nanovials in a tube where they bind to antibodies on the nanovials for cell surface markers (CD27 or CD45). The loaded nanovials are incubated to accumulate secreted IgG on the surface via anti-IgG capture antibodies. Nanovials are then stained with fluorescent or oligo-barcoded anti-IgG, as well as viability dyes and other surface marker stains. Stained nanovials and associated cells are analyzed by flow cytometry (LSR II flow cytometer), imaging flow cytometry (ImageStream), or sorted (Nanocellect WOLF) for single-cell transcriptomics using the 10X Chromium system. Data linking IgG secretion with surface markers/functional dyes and transcriptomes at the single-cell level is acquired and analyzed.

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