Ivis lumina k

Six-week old athymic nude female mice were purchased from Charles River Laboratories. In the intracardiac injection metastasis model, mice were injected with A549 cells expressing luciferase under anesthesia, as previously described¹⁵ . Briefly, 1cc syringe with a 28 gauge needle (BD, #329410) was loaded with 5 × 10⁵ cells in 100 μL PBS. Then the needle was inserted into the left ventricle of the heart through the second intercostal space, followed by injection of the cells at a very low pace once trace of blood was pumped into the syringe. Following injection, the animal was placed in a clean cage with a heating pad until full recovery. The success of intracardiac injection was further confirmed by in vivo bioluminescence imaging immediately after injection (IVIS® Lumina K, PerkinElmer). For MK-2206 experiments, a total of 27 mice were randomly divided into 3 treatment groups: vehicle (30% Captisol), MK-2206 (60 mg/kg), and MK-2206 (120 mg/kg), administered via oral gavage three times per week. Treatments were started on the day after intracardiac injection, and were for two weeks. Mice were monitored by in vivo bioluminescence imaging once weekly. All animal experiments were conducted under a protocol approved by the Georgetown University Animal Care Committee.

Quantum dots (QDs) are inorganic probes comprising CdSe/ZnS-core/shell semiconductor nanocrystals, and a useful low-cytotoxicity tool for fluorescent cell labeling. The transduction of QDs was conducted as previously described (Yukawa et al., 2009 (link)). Briefly, A549 and PC-9 cells were incubated in a culture medium containing 2 nM Qdot™ 800 ITK™ Carboxyl Quantum Dots (Invitrogen) for 24 h. After washing with PBS, a suspension of 2.5 × 10⁶ QD-labeled lung cancer cells was injected directly into each part of a decellularized rat lung scaffold using 30G needles. Following overnight perfusion cultivation (1 mL/min), the rat lung scaffold was subjected to IVIS Imaging System, IVIS Lumina K (PerkinElmer). The fluorescence derived from QDs800 in specimen was detected with IVIS Lumina K at the condition of excitation filter: 740 nm ± 10 nm, emission filter: 790 nm ± 15 nm.

For in vivo luminescence imaging, HeLa tumor-bearing mice were intravenously injected with CP1-NCs. The luminescence signals of the mice were collected (λ_ex = 690 nm) through an IVIS Lumina K in vivo imaging system (PerkinElmer) using a xenon lamp which was equipped with different long- and band-pass filters. An Andor EMCCD-DU897 camera was used as an imaging detector. Images were taken at 2, 4, 8, 12, and 24 h postinjection. After 24 h, the tumor and major organs were collected and the fluorescence intensity was analyzed to confirm the accurate therapeutic time.