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Sunnybrook Research Institute

10 ARTICLES PUBLISHED IN JoVE

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Medicine

MRI-guided Disruption of the Blood-brain Barrier using Transcranial Focused Ultrasound in a Rat Model
Meaghan A. O'Reilly 1, Adam C. Waspe 1,2, Rajiv Chopra 1,2, Kullervo Hynynen 1,3
1Imaging Research, Sunnybrook Research Institute, 2Department of Medical Biophysics, University of Toronto, 3Department of Medical Biophysics, and Institute of Biomaterials & Biomedical Engineering (IBBME), University of Toronto

Microbubble-mediated focused ultrasound disruption of the blood-brain barrier (BBB) is a promising technique for non-invasive targeted drug delivery in the brain1-3. This protocol outlines the experimental procedure for MRI-guided transcranial BBB disruption in a rat model.

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Medicine

Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging
Jill J. Weyers 1, Dara D. Carlson 1, Charles E. Murry 1,2, Stephen M. Schwartz 1, William M. Mahoney, Jr. 1
1Department of Pathology, Center for Cardiovascular Biology, and Institute for Stem Cell and Regenerative Medicine, University of Washington, 2Departments of Bioengineering and Medicine/Cardiology, University of Washington

Visualization of the coronary vessels is critical to advancing our understanding of cardiovascular diseases. Here we describe a method for perfusing murine coronary vasculature with a radiopaque silicone rubber (Microfil), in preparation for micro-Computed Tomography (μCT) imaging.

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Medicine

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)
Aida Gordanpour 1, Robert K. Nam 2, Linda Sugar 3, Stephanie Bacopulos 1, Arun Seth 1,3,4
1Department of Laboratory Medicine & Pathobiology, University of Toronto, 2Division of Urology, Sunnybrook Health Sciences Centre, Toronto, Canada, 3Department of Anatomic Pathology, Sunnybrook Health Sciences Centre, Toronto, Canada, 4Biological Sciences, Sunnybrook Research Institute

Quantitative Real Time polymerase chain reaction (qPCR) is a rapid and sensitive method to investigate the expression levels of various microRNA (miRNA) molecules in tumor samples. Using this method expression of hundreds of different miRNA molecules can be amplified, quantified, and analyzed from the same cDNA template.

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Biology

Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands
Rushin Shojaii 1, Stephanie Bacopulos 2,3, Wenyi Yang 2,3, Tigran Karavardanyan 4, Demetri Spyropoulos 5, Afshin Raouf 6, Anne Martel 1,4, Arun Seth 2,3
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.

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Medicine

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy
Amanda Tracz 1, Michalis Mastri 1, Christina R. Lee 2, Roberto Pili 1, John M. L. Ebos 1
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.

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Neuroscience

Long-term Time Lapse Imaging of Mouse Cochlear Explants
Joanna F. Mulvaney 1, Alain Dabdoub 1,2,3
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.

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Developmental Biology

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound
Janet M. Denbeigh 1,2, Brian A. Nixon 1,2, Mira C. Puri 1,2,3, F. Stuart Foster 1,2
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.

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Medicine

MRI-guided Focused Ultrasound Thalamotomy for Patients with Medically-refractory Essential Tremor
Ying Meng 1, Yuexi Huang 2, Benjamin Solomon 2, Kullervo Hynynen 2, Nadia Scantlebury 1, Michael L. Schwartz 1, Nir Lipsman 1
1Division of Neurosurgery, Sunnybrook Health Sciences Centre, 2Sunnybrook Research Institute, Sunnybrook Health Sciences Centre

High-intensity MRI guided focused ultrasound is an emerging noninvasive technique to precisely ablate brain tissue. It has been shown to be safe and effective in treating medically-refractory essential tremor. This article describes the protocol for thalamotomy from patient selection to equipment setup to post-treatment follow-up.

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Bioengineering

Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability
Charissa Poon 1,2, Melina Mühlenpfordt *3, Marieke Olsman *3, Spiros Kotopoulis 4,5, Catharina de Lange Davies 3, Kullervo Hynynen 1,2,6
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.

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Advanced Techniques for Studying Blood Flow
Jill J. Weyers 1,2, Nilesh R. Ghugre 1,3,4
1Physical Sciences Platform, Sunnybrook Research Institute, 2Institute of Biomedical Engineering, University of Toronto, 3Schulich Heart Research Program, Sunnybrook Research Institute, 4Department of Medical Biophysics, University of Toronto

Advanced Techniques for Studying Blood Flow

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