Ivis lumina xr 3 in vivo imaging system

Subcutaneous injection of murine 4T1 cells into Balb/c mice led to the formation of primary tumors. At 21 days after 4T1 cell injection, when the tumor diameter was >7 mm (range, 7–10 mm), ten mice were divided into two groups: treated with LnNP@ZIF8 (LnNP@ZIF8) or treated with LnNP@ZIF8-DOX (LnNP@ZIF8-DOX). The mice were anesthetized using inhaled isoflurane (RWD Life Science, Shenzhen, China). The tumor site was sanitized with a 75% ethanol tincture before treatment. All the procedures were performed aseptically. Then, a 100µL mixture of the LnNPs in PBS (5 mg/mL) was injected subcutaneously into the 4T1 tumors.
Eight of ten SCID tissue-derived orthotopic mice models constructed successfully. Then, a 100 µL mixture of the LnNP@ZIF8-DOX (5 mg/mL) in PBS was injected subcutaneously into the MCF-7 WT and MCF-7 ADR tumors.
NIR-II fluorescent images were recorded at 24 h after probe injection using an in vivo imaging system equipped with an IVIS Lumina XR III in vivo imaging system (PerkinElmer, Waltham, MA, USA).
All mice were sacrificed by cervical dislocation after CO₂ euthanasia.

Animal experiments were operated according to the Guidelines for the Care and Use of Laboratory Animals published by the National Institutes of Health [26 (link)] and approved by the Ethics Committee of Shengjing Hospital of China Medical University. To establish a PTC lung metastasis model, thymic BALB/c nude mice (16–18 g) were used. Stable TPC-1 cells expressing a luciferase and shRNA for circ_102002 (sh-circ_102002) or the control cells (sh-NC) were used. Mice (5 mice/group) were intravenously injected with these cells (1 × 10⁶). At 21 days post-implantation, the tumor-bearing mice were intraperitoneally injected with 100 μl of d-luciferin (10 mg/ml in PBS, Promega, Madison, WI, USA). After 10 min, the fluorescence images of the tumor and lung metastasis in the mice were imaged under anesthesia with 2.5% isofluorane using an IVIS Lumina XR III in vivo imaging system (PerkinElmer, Waltham, CA). Luminescence was expressed as photons/s per region of interest minus background luminescence for a similarly sized region. The lungs were dissected from mice and photographed. Additionally, haematoxylin and eosin (H&E) staining and IHC analysis of HAS2, p-FAK, p-Akt, E-cadherin, and N-cadherin in the tumor tissues were, respectively, conducted. All the antibody information was listed in Supplementary Table S2.