Under an approved IACUC protocol (ND #14-060), six-week-old female nude mice nu/nu (Charles River, Wilmington, MA) were maintained at the Freimann Life Science Center (University of Notre Dame). In the pilot study (4 mice per group), 2×106 SKOV3IP-RFP, SKOV3IP-ARID3BFL, and SKOV3IP-ARID3BSH cells in 300 μl of phosphate buffered saline (PBS; 137 mM NaCl, 2.7 mM KCl, and 11.9 mM phosphate buffer, pH 7.4) were injected intraperitoneally (IP) into nude mice. Mice had visible tumors after 3 weeks and were euthanized. For the survival studies and analysis of tumor growth, 1×106 SKOV3IP-RFP (9 mice), SKOV3IP-ARID3BFL (16 mice), and SKOV3IP-ARID3BSH (14 mice) cells in 300 μl of PBS cells were each injected into nude mice. Two separate experiments were conducted on independent transductions of the cells. Mice were imaged weekly. When the mice showed signs of illness they were imaged twice weekly and then euthanized. Ascites fluid was collected using a 25 G needle and 5 mL syringe. If no ascites fluid was present a peritoneal wash was performed by injecting 2 mL of PBS IP and then collecting the fluid. Fluorescent imaging of tumor growth in live animals were acquired at the Notre Dame Integrated Imaging Facility using the Multispectral FX (Carestream, Rochester, NY
Multispectral fx
Manufactured by Carestream
Sourced in Canada, United States
The Multispectral FX is a lab equipment product that captures images using multiple wavelengths of light. It provides high-quality, multiband imaging capabilities for various applications.
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Lab products found in correlation
3 protocols using multispectral fx
1
Tracking Ovarian Tumor Growth in Nude Mice
2
Quantifying Bone and Cartilage Formation in Distraction Osteogenesis
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Digital X‐ray images were taken at days 13 and 27 after surgery, using a Kodak Multispectral FX instrument (Carestream Health, Toronto, Canada). The distraction gap was outlined from the outside corners of the two proximal and the two distal cortices, forming a quadrilateral region of interest (ROI). The X‐ray density of newly formed bone was analysed using Bruker MI SE software, v. 7.1.3 (Scrum, Osaka, Japan). After the animals had been sacrificed, the distracted tibias were fixed in 10% neutral buffered formalin for 24 h at room temperature. Samples were decalcified with Morse's solution (10% sodium citrate, 20% formic acid) overnight at 4 °C. The samples were then washed in PBS and embedded with OCTTM (Sakura Finetek, Tokyo, Japan), after which 8 µm sections were made on a cryostat (Leica CM 3050S, Leica Microsystems, Wetzlar, Germany). The cryosections were stained with haematoxylin and eosin (H&E) and safranin O–fast green. In the sections of each sample, the percentage area of newly formed cartilage and bone was measured. The pixel number of each area in the distraction zone was counted using Image J, v. 1.43 (NIH, Bethesda, MD, USA) (see supporting information, Figure S1 B).
3
Ovarian Cancer Xenograft Survival Study
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Under an approved IACUC (Institutional Animal Care and Use Committee) protocol (ND #14-060), 6-week-old female nude mice nu/nu (Charles River, Wilmington, MA, USA) were maintained at the Freimann Life Science Center (University of Notre Dame). For the survival studies and analysis of tumor growth, 1 × 106 Skov3IP-RFP+shControl (10 mice), Skov3IP-ARID3B+shControl (10 mice), and Skov3IP-ARID3B+shPROM1 (10 mice) cells in 200 μL of PBS were each injected intraperitoneally into nude mice. Starting 14 days after injection, mice were imaged weekly at the Notre Dame Integrated Imaging Facility using the Multispectral FX (Carestream, Rochester, NY, USA).
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