Fig. 2: Combined tyramide-barcode and conjugated primary antibody staining in tonsil.
From: Tyramide-conjugated DNA barcodes enable signal amplification for multiparametric CODEX imaging
CD3e (1:100 dilution) and CD20 (1:200 dilution) were stained using commercial barcode-conjugated primary antibodies (Akoya Biosciences), and staining of CD4 and CD163 was performed using tyramide-barcodes with mouse anti-CD4 (1:100 dilution, clone 4B12, Leica Biosystems) and mouse anti-CD163 (ready-to-use format, clone 10D6, Leica Biosystems) primary antibodies. The composite image is colored as follows: DAPI = blue, CD20 = red, CD4 = yellow, CD3e = green, and CD163 = magenta. The Keyence BZ-X800 microscope used 100% excitation intensity, High Resolution imaging mode, and a Nikon 20x PlanApo lambda 0.75 NA objective. Background subtraction and image stitching were performed using Akoya’s CODEX Processor software.
