Figure 1.

Analysis of bone marrow (BM) trephines using digital spatial profiling (DSP).

(A) Representative regions of interest (ROIs) from each sample in the study. Using fluorescent immunohistochemistry [CD3 (green), CD45 (red) and nuclear stain Syto83 (blue)], 300-μm ROIs were selected in each tissue, and the expression levels of 31 proteins within each were determined by staining with nCounter barcode-tagged antibodies. (B) Raw DSP counts in the normal BM trephine. ROIs were placed within the tissue (black), decalcified trabeculum (blue) and blank slide (red) to determine the background level of staining for each antigen. (C) Correlation between ROI mean fluorescence and normalised DSP counts in the normal BM trephine. Neg. cont., negative control; Beta-2-M, beta-2-microglobulin; Beta-Cat, beta-catenin; GZMB, granzyme B; M IgG2a, mouse IgG2a; Pan-CK, pan-cytokeratin.