10–14 days prior to the experiment, 6-week-old female CD1-nu mice were inoculated with 5 × 106 A431-CCK2R cells (0.1 mL, 5 × 107 cells/mL in PBS) subcutaneously in both shoulders. The mice were randomly assigned to groups of two animals. On the day of the experiment, 177Lu-labeled conjugates (200 pmol, 100 µL, 2 µM in PBS, 20 MBq, 20 µg/kg) were injected via the tail vein. The mice were kept with food and water ad libitum. SPECT scans of 40 min (1 and 4 h p.i., frame duration 60 sec) or 60 min (24 h p.i., frame duration 80 sec) were acquired after an initial CT scan of 7.5 min (CT tube voltage 55 kV, CT tube current 145 µA) with a multi-pinhole NanoSPECT/CTplus (Bioscan Inc., France, and Mediso, Hungary). For the duration of the SPECT/CT scans, mice were kept under anesthesia with a mixture of isoflurane (1.5–2.5%) and oxygen and warmed by a constant air flow of 37 °C. The images were reconstructed with HiSPECT and analyzed in VivoQuant. A post-reconstruction filter was applied (Gauss, full width at half max, 1 mm) to the SPECT images, and the scale was adjusted to visualize important organs and tissues (0.001–0.004 kBq/voxel). n = 2 animals per group.

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