Xenogen ivis spectrum in vivo imaging system

All animals were treated in accordance with NIH Guidelines for the Care and Use of Laboratory Animals and the established Institutional Animal Use and Care protocol at the University of Illinois, Chicago. Xenograft studies utilized NCr nu/nu athymic female mice 6-8 weeks in age (Taconic). Mice were housed in a temperature and light-controlled environment (12 hours light and 12 hours dark) and provided food and water ad libitum. For xenograft experiments, OVCAR8-RFP cells (5 x 10⁶) were injected intraperitoneally (IP) per mouse and tumor growth was monitored using Xenogen IVIS® Spectrum In Vivo Imaging System (PerkinElmer) as previously described (20 ). Once all the mice formed tumors (~4 weeks), the mice were separated into 2 treatment groups and dosed once every two days with 0.5 mg/kg of verticillin A encapsulated nanoparticles (eNP-VA) and empty nanoparticles (eNP) for a total of 12 days. Mice were IVIS imaged twice weekly (535 nm excitation, and 620 nm emission, Exposure time: 2 seconds, F stop: 2). Living Image 4.0 software was used to quantify the average abdominal radiant efficiency and normalization was performed using Day 0 radiant efficiency. At week 7, all animals were sacrificed and tumors were collected for histological analysis.

5 × 10⁶ OVCAR8-RFP cells were xenografted IP per mouse and tumor growth was monitored with the Xenogen IVIS® Spectrum In Vivo Imaging System (PerkinElmer) as previously described (47 (link)). When tumors became detectable via IVIS® imaging, mice were separated into treatment groups of 5–6 mice and dosed once daily 3 times per week with paclitaxel (5 mg/kg), PHY34 (0.75 mg/kg), or vehicle (10% DMSO:40% PEG300:50% H₂O) for 3 weeks. Mice were weighed 3 times a week and imaged once weekly with an exposure of 2 seconds, an Fstop of 2, and an excitation and emission wavelength of 535 and 620 nm, respectively. Average abdominal radiant efficiency was quantified with the system’s Living Image 4.0 software and normalized to Day 0. After week 3, mice were sacrificed and tumors were collected for immunohistochemistry. Statistics were generated with two-way ANOVA with Dunnett’s multiple comparisons to vehicle control.

The Xenolight Rediject Inflammation Probe (PerkinElmer, Waltham, MA) is an intravital luminescent dye that penetrates phagocytic cells and enables visualization of joint infiltration. Probe was administered to mice 7 days after arthritis induction by i.p. injection according to the manufacturer’s instructions. Joint inflammation was quantified using the Xenogen IVIS Spectrum in vivo Imaging System (Perkin Elmer).