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Representative in vivo serial fluorescence images and region of interest (ROI) analysis of singlet domain antibodies (VHH) singlet (a) and iron oxide nanoparticlePEG2000/750 (IONPPEG2000/750) VHH triplet (b). ROIs were drawn to track the fluorescence signal change of major ROIs including kidney and brain (prone position), left front paw, and liver (supine image) over time. Scale bar represents near-infrared (NIR) image intensity measured by Pearl. (a) In vivo serial fluorescence prone and supine position images before and 2 min, 1 h, 2 h, 4 h, 8 h, 1 day, and 2 days after IV bolus injection of VHH singlet. (b) In vivo serial fluorescence prone and supine position images before and 2 min, 1 h, 2 h, 4 h, 8 h, 1 day, and 2 days after IV bolus injection of IONPPEG2000/750 VHH triplet. 1e1e36bf2d