Programed death ligand-1 (PD-L1) expression is currently the only predictive biomarker for cancer immunotherapy. Since PD-L1 expression in tumors is largely heterogeneous, in-vivo detection and quantification of PD-L1 in intact tumors is of major interest. Here we employ fluorescence lifetime (FLT) imaging for in-vivo detection and quantification of PD-L1 expression using an anti-PD-L1 antibody conjugated to IRDye800CW (αPDL1-800). We show that FLT imaging accurately identifies heterogeneous PD-L1 expression in tumors. Tumor areas of high PD-L1 levels were spatially correlated to significantly longer FLTs of αPDL1-800 and the distribution of PD-L1 in deep-seated (>1cm depth) tumors was achieved using FLT tomography.
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