Ccd camera

TICs within the two passages subsequent to lentiviral transduction were harvested with accutase and resuspended in serum-free supplemented-medium. Three hundred thousand cells were injected into the pons of adult mice in a total volume of 3 μL (i.e. 100,000 cells/μl) as described above. Mice were kept under specific pathogen-free conditions in air-filtered cages and received food and water ad libitum. Tumor growth was monitored using an IVIS 50 system with charge-coupled device (CCD) camera (PerkinElmer) every 2-3 weeks. Mice were anesthetized as described previously and imaged 10 min after intra-peritoneal injection of luciferin (150 mg/kg body weight, Promega). Radiance signal intensity (normalized photon counts in photons/second/cm²/steradian) in the pons region was quantified using Living Image software (Perkin Elmer). Doubling time of the CDOX models was estimated based on bioluminescence exponential curves.

Mice were anesthetized with an intraperitoneal injection of 75 mg/kg of sodium pentobarbital (P276000, Toronto Research Chemicals) and transferred to an imaging chamber. Mice were imaged from dorsal or ventral view using Xenogen IVIS Lumina II system equipped with the charge-coupled device (CCD) camera (PerkinElmer, MA). Bioluminescence images were quantified using the Xenogen Corperation Living Image software (PerkinElmer, MA). The total intensity was calculated from a user-defined area of interest covering the whole animal body except the paws. Lower limit of quantification of imaging is approximately 1×10⁸ photons (p).

The luciferase substrate; luciferin (150 mg/kg, D-Luciferin potassium salt, Xenogen, Gold Biotechnology, St. Louis, MO, USA) was inoculated intraperitoneally (i.p.) into BALB/c mice 10 min before bioluminescent signals were acquired. After 7 min of injection, the mice were anaesthetized by inhalation with 3% isoflurane in oxygen (v/v) at a flow rate of 2.5 L/min until no movement was seen (3 min approx.). Mice were then imaged using the IVIS Spectrum in vivo Imaging System (PerkinElmer, Waltham, MA, USA), and the images were acquired with the Living Image software (version 4). Emitted photons were gathered by a charge couple device (CCD) camera (PerkinElmer, USA) using the medium resolution (medium binning) mode. A circular region of interest (ROI) encompassing the nodular area on the rump was drawn to quantify the bioluminescence, expressed as radiance in numbers of photons/sec [36 (link)].