Sunnybrook Research Institute

10 ARTICLES PUBLISHED IN JoVE

Medicine

MRI-guided Disruption of the Blood-brain Barrier using Transcranial Focused Ultrasound in a Rat Model

Meaghan A. O'Reilly¹, 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 brain^1-3. This protocol outlines the experimental procedure for MRI-guided transcranial BBB disruption in a rat model.

Medicine

Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging

Jill J. Weyers¹, Dara D. Carlson¹, Charles E. Murry^1,2, Stephen M. Schwartz¹, 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.

Medicine

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)

Aida Gordanpour¹, Robert K. Nam², Linda Sugar³, Stephanie Bacopulos¹, 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.

Biology

Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands

Rushin Shojaii¹, Stephanie Bacopulos^2,3, Wenyi Yang^2,3, Tigran Karavardanyan⁴, Demetri Spyropoulos⁵, Afshin Raouf⁶, 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.

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.

Neuroscience

Long-term Time Lapse Imaging of Mouse Cochlear Explants

Joanna F. Mulvaney¹, 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.

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.

Medicine

MRI-guided Focused Ultrasound Thalamotomy for Patients with Medically-refractory Essential Tremor

Ying Meng¹, Yuexi Huang², Benjamin Solomon², Kullervo Hynynen², Nadia Scantlebury¹, Michael L. Schwartz¹, Nir Lipsman¹

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.

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 Davies³, 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.

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