Ivis lumina series 3 imaging system

Mice were handled in accordance with institutional (SCXK(jing) 2017-0005)NIFDC, Beijing, China) guidelines for laboratory animal care and use, and the Animal Care and Use Committee at the NIFDC approved the study protocol. Four to five-week-old female BALB/c mice (n = 5 per group) were obtained from the Institute of Laboratory Animal Resources of NIFDC. To challenge the mice, 4.5 × 10⁶ PFU HAdV5 were administered via the tail vein to groups of five mice. Cardamomin (CDM) was dissolved in 0.5% methylcellulose (Sigma-Aldrich, St. Louis, MO) and aliquoted into three doses (150 mg/kg, 75 mg/kg, and 37.5 mg/kg), which were IP-injected into three groups of mice. The drug was administered 6 h before infection, then once daily until 5 dpi. Bioluminescence imaging (BLI) analysis was performed with the IVIS Lumina Series III Imaging System (PerkinElmer, Baltimore, MD) as previously described (Zhang et al., 2017 (link)). We used Living Image software (Caliper Life Sciences, Baltimore, MD) to analyze the regions of interest, which were represented by the total flux in photons per second.

All animal experiments were approved by the Tel-Aviv Sourasky Medical Center (TASMC) ethics committee (see Ethics, below). The experimental mice were 10–12 weeks old NOD/SCID/IL2Rγ^−/− obtained from The Jackson Laboratories (Bar Harbor, ME, USA). They were maintained in a specific pathogen-free facility of the TASMC. A total of 2 × 10⁶ NAR-LUC tumor cells were injected intraperitoneally. After 18–23 days, ErbB2CAR or UNT T cells were administered to tumor-bearing mice by either IP or IV injection. The anti-tumor response was evaluated by following survival up to 92 days and by in vivo imaging detection of luciferase signals expressed by NAR LUC cells. Bio-luminescence imaging was performed following IP injection of luciferin (375 μg/kg) with the animal anaesthetized with ketamine (100 mg/kg) and xylazine (20 mg/kg) in accordance with institutional guidelines. The mice were monitored by an IVIS Lumina series III imaging system (Perkin Elmer, Waltham, MA, USA). For flow cytometric analysis, the mice were euthanized either after a 10% weight loss or 45 days from the initiation of the experiment.

Metastasis development was monitored by in vivo bioluminescence imaging (IVIS Lumina Series III Imaging system, PerkinElmer) on day 1 (baseline imaging), day 14 and day 28 after tumor cell injection. For image acquisition the mice received an intraperitoneal injection of Luciferin (Luciferin substrate cat. no.: 5306500001, Calbiochem; dosage: 150 mg/kg; stock solution: 30 mg/mL in H₂O; application volume: 100–150 µL). After 3 min of incubation the animals were sedated with 5% isoflurane and then transferred to the imaging chamber with 2% isoflurane and 37 °C. Imaging was started 10 min after Luciferin injection using the XFOV-24 lense and an exposure time of 180 sec (medium bining, 1.2 F/Stop, minimum target count luminescent: 10,000). Images were taken from the ventral as well as from the dorsal view.