Royal Prince Alfred Hospital View Institution's Website 6 articles published in JoVE Biology The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury Jully Gogoi-Tiwari1, Julia Köhn-Gaone1, Corey Giles2, Dirk Schmidt-Arras3, Francis D. Gratte1,4, Caryn L. Elsegood1, Geoffrey W. McCaughan5,6,7, Grant A. Ramm8,9, John K. Olynyk10,11, Janina E.E. Tirnitz-Parker1,12 1School of Biomedical Sciences & Curtin Health Innovation Research Institute, Curtin University, 2School of Public Health & Curtin Health Innovation Research Institute, Curtin University, 3Institute of Biochemistry, Christian-Albrechts-University, 4School of Veterinary and Life Sciences, Murdoch University, 5Centenary Institute of Cancer Medicine and Cell Biology, The University of Sydney, 6Royal Prince Alfred Hospital, 7A.W. Morrow Gastroenterology and Liver Centre, 8QIMR Berghofer Medical Research Institute, 9Faculty of Medicine and Biomedical Sciences, The University of Queensland, 10Fiona Stanley and Fremantle Hospitals, 11School of Medical and Health Sciences, Edith Cowan University, 12School of Medicine and Pharmacology, University of Western Australia Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses. Medicine Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses Chi Ching Goh1,2, Jackson LiangYao Li1, David Becker3, Wolfgang Weninger4,5,6, Veronique Angeli2,7, Lai Guan Ng1,2,5,8 1Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, 2Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 3Lee Kong Chian School of Medicine, Nanyang Technological University, 4Centenary Institute for Cancer Medicine and Cell Biology, 5Discipline of Dermatology, University of Sydney, 6Department of Dermatology, Royal Prince Alfred Hospital, 7LSI Immunology Programme, National University of Singapore, 8School of Biological Sciences, Nanyang Technological University This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury. Medicine Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids Kimberley A. Beaumont1,2, Andrea Anfosso1,2, Farzana Ahmed3, Wolfgang Weninger*1,4,5, Nikolas K. Haass*1,3,5 1The Centenary Institute, 2Sydney Medical School, University of Sydney, 3The University of Queensland Diamantina Institute, Translational Research Institute, The University of Queensland, 4Department of Dermatology, Royal Prince Alfred Hospital, 5Discipline of Dermatology, University of Sydney We describe two complementary methods using the fluorescence ubiquitination cell cycle indicator (FUCCI) and image analysis or flow cytometry to identify and isolate cells in the inner G1 arrested and outer proliferating regions of 3D spheroids. Biology Isolation of Small Noncoding RNAs from Human Serum Samantha Khoury1, Pamela Ajuyah1, Nham Tran2,3 1School of Medical and Molecular Biosciences, Faculty of Science, University of Technology, Sydney, 2Centre for Health Technologies, Faculty of Engineering and Information Technology, University of Technology, Sydney, 3The Sydney Head and Neck Cancer Institute, Sydney Cancer Centre, Royal Prince Alfred Hospital This protocol describes a method for extracting small RNAs from human serum. We have used this method to isolate microRNAs from cancer serum for use in DNA arrays and also singleplex quantitative PCR. The protocol utilizes phenol and guanidinium thiocyanate reagents with modifications to yield high quality RNA. Medicine Murine Model of Wound Healing Louise Dunn1,2, Hamish C. G Prosser1,2, Joanne T. M. Tan1,2, Laura Z. Vanags1,2, Martin K. C. Ng1,2,3, Christina A. Bursill1,2 1The Heart Research Institute, 2Sydney Medical School, University of Sydney, 3Cardiology Department, Royal Prince Alfred Hospital A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings. Immunology and Infection Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing Jerry Zhou1, Larissa Belov1, Michael J. Solomon2, Charles Chan3, Stephen J. Clarke4, Richard I. Christopherson1 1School of Molecular Bioscience, University of Sydney, 2Department of Surgery, Royal Prince Alfred Hospital, 3Department of Anatomical Pathology, Department of Anatomical Pathology, 4Department of Medicine, Concord Repatriation General Hospital We described a procedure for the disaggregation of colorectal cancer (CRC) to produce viable single cells, which are then captured on customized antibody microarrays recognizing surface antigens (DotScan CRC microarray). Sub-populations of cells bound to the microarray can be profiled by fluorescence multiplexing using monoclonal antibodies tagged with fluorescent dyes.
