Extended Data Fig. 4: Example of 27-plex CyCIF experiment with tumour sample and tonsil control tissues.
From: Temporal and spatial topography of cell proliferation in cancer
a. Example of positive and negative staining for all markers in CyCIF experiment through rounds of cyclic imaging. Three independent samples are shown: Ctrl is a non-malignant tonsil tissue sample (‘control’), #1 and #2 are glioma samples. Plots are single-cell kernel smoothing (KS) density estimation for patient samples from respective images (median per pixels within the cell area, log2 FAU, not normalized, black = tonsil, yellow = glioma sample #1, magenta = glioma sample #2, n = 4,278, 2,629, and 2,609 cells, respectively). Each row of images and data is a successive round of CyCIF acquired from the same tissue area (Rx is the xth round of imaging). All images from antibody channels were linearly contrasted between 0 and 2000 fluorescence units for ease of comparison. Scale bar, 50 µm. Examples from tissue microarray containing 176 independent glioma specimens and 8 independent control specimens. b. Scatter plots of the single-cell correlation of the signal intensity of unconjugated antibodies to the indicated markers versus their fluorophore conjugated versions from 142 samples of tissue from 75 patients (Pantomics TMA BRC15010). Pearson correlation coefficients (C) are shown. All correlation had p-value < 10-175. c. Plot of the phospho-Rb signal from MPI0, MPI + 1/Ki-67-, and MPI + 1/Ki-67+ cells from Pantomics TMA BRC15010 acquired using both conjugated (‘direct IF’) and unconjugated (‘indirect IF’) phospho-Rb antibodies (n = 74 cores, Line is mean value).
