Figure 3

CD298 and b2m enable robust live cell barcoding of heterogeneous samples. (A) Schematic representation of the barcoding procedure. Individual samples are barcoded with a combination of three different b2m and CD298 antibodies, pooled, stained and acquired. Samples can then be debarcoded using the supplied barcoding matrix. Arrows point out sample 10 and 13 which are used as examples in A and C. (B) 20 PBMC samples were barcoded using the above described method. Shown are live, single cells from combined samples before debarcoding. (C) Combined samples can be debarcoded using existing algorithms and software. (D) Example of a debarcoded sample positive for three of the available six barcodes. (E) Signal intensities of all barcode isotopes from combined anti-b2m and anti-CD298 antibodies on debarcoded samples as in E. (F) PBMC and HeLa samples were barcoded, combined and stained with anti-CD45 (left). Frequency of CD45⁺ in debarcoded cells (right). (G) Exemplary biaxial plots demonstrating CD45 staining in debarcoded cells as in F, assigned to a PBMC (left) or HeLa sample (right).