Sunnybrook Research Institute 5 articles published in JoVE Bioengineering Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability Charissa Poon1,2, Melina Mühlenpfordt*3, Marieke Olsman*3, Spiros Kotopoulis4,5, Catharina de Lange Davies3, Kullervo Hynynen1,2,6 1Physical Sciences Platform, Sunnybrook Research Institute, 2Institute of Biomedical Engineering, University of Toronto, 3Department of Physics, Norwegian University of Science and Technology, 4Department of Clinical Medicine, University of Bergen, 5Exact Therapeutics AS, 6Department of Medical Biophysics, University of Toronto This protocol describes the surgical and technical procedures that enable real-time in vivo multiphoton fluorescence imaging of the rodent brain during focused ultrasound and microbubble treatments to increase blood-brain barrier permeability. Developmental Biology Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound Janet M. Denbeigh1,2, Brian A. Nixon1,2, Mira C. Puri1,2,3, F. Stuart Foster1,2 1Department of Medical Biophysics, University of Toronto, 2Sunnybrook Research Institute, 3Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto Here, we present a protocol to inject ultrasound microbubble contrast agents into living, isolated late-gestation stage murine embryos. This method enables the study of perfusion parameters and of vascular molecular markers within the embryo using contrast-enhanced high-frequency ultrasound imaging. Neuroscience Long-term Time Lapse Imaging of Mouse Cochlear Explants Joanna F. Mulvaney1, Alain Dabdoub1,2,3 1Biological Sciences Platform, Sunnybrook Research Institute, 2Department of Otolaryngology - Head and Neck Surgery, University of Toronto, 3Department of Laboratory Medicine and Pathobiology, University of Toronto Live imaging of the embryonic mammalian cochlea is challenging because the developmental processes at hand operate on a temporal gradient over ten days. Here we present a method for culturing and then imaging embryonic cochlear explant tissue taken from a fluorescent reporter mouse over five days. Biology Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands Rushin Shojaii1, Stephanie Bacopulos2,3, Wenyi Yang2,3, Tigran Karavardanyan4, Demetri Spyropoulos5, Afshin Raouf6, Anne Martel1,4, Arun Seth2,3 1Department of Medical Biophysics, University of Toronto, 2Platform Biological Sciences, Sunnybrook Research Institute, 3Department of Laboratory Medicine and Pathobiology, University of Toronto, 4Physical Sciences, Sunnybrook Research Institute, 5Department of Pathology and Laboratory Medicine, Medical University of South Carolina, 6Manitoba Institute of Cell Biology, University of Manitoba We present an image registration approach for 3-dimensional (3D) histology volume reconstruction, which facilitates the study of the changes of an organ at the level of macrostructures made up of cells . Using this approach, we studied the 3D changes between wild-type and Igfbp7-null mammary glands. Medicine Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy Amanda Tracz1, Michalis Mastri1, Christina R. Lee2, Roberto Pili1, John M. L. Ebos1 1Genitourinary Section, Department of Medicine, Roswell Park Cancer Institute, 2Biological Sciences Platform, Sunnybrook Research Institute Models of spontaneous metastatic renal cell carcinoma (RCC) disease progression can be used for evaluating treatments in a clinically relevant setting. This protocol demonstrates different procedures for orthotopic kidney tumor cell implantation, proper nephrectomy, and finally outlines a necropsy guide for visual and bioluminescent scoring of metastatic burden and localization.
