Monash Health 4 articles published in JoVE Immunology and Infection Supervised Machine Learning for Semi-Quantification of Extracellular DNA in Glomerulonephritis Kim Maree O'Sullivan1, Sarah Creed2, Poh-Yi Gan1, Stephen R. Holdsworth1,3 1Department of Medicine, Monash University, 2Monash Micro Imaging, Monash University, 3Immunology Department, Monash Health Extracellular DNA (ecDNA) released during cell death is proinflammatory and contributes to inflammation. Measurement of ecDNA at the site of injury can determine the efficacy of therapeutic treatment in the target organ. This protocol describes the use of a machine learning tool to automate measurement of ecDNA in kidney tissue. Behavior Radiotracer Administration for High Temporal Resolution Positron Emission Tomography of the Human Brain: Application to FDG-fPET Sharna D. Jamadar1,2,3, Phillip G.D. Ward1,2,3, Alexandra Carey1,4, Richard McIntyre1,4, Linden Parkes1,3, Disha Sasan1, John Fallon1, Edwina Orchard1,2,3, Shenpeng Li1,5, Zhaolin Chen1,5, Gary F Egan1,2,3 1Monash Biomedical Imaging, Monash University, 2Australian Research Council Centre of Excellence for Integrative Brain Function, 3Turner Institute for Brain and Mental Health, Monash University, 4Department of Medical Imaging, Monash Health, 5Department of Electrical and Computer Systems Engineering, Monash University This manuscript describes two radiotracer administration protocols for FDG-PET (constant infusion and bolus plus infusion) and compares them to bolus administration. Temporal resolutions of 16 s are achievable using these protocols. Medicine Optical Clearing and Imaging of Immunolabeled Kidney Tissue Turgay Saritas1, Victor G. Puelles1,2,3, Xiao-Tong Su4, David H. Ellison4,5,6, Rafael Kramann1,7 1Division of Nephrology and Clinical Immunology, University Hospital RWTH Aachen, 2III. Department of Medicine, University Medical Center, Hamburg-Eppendorf, 3Department of Nephrology, Monash Health, 4Division of Nephrology and Hypertension, Oregon Health and Science University, 5Renal Section, Veterans Affairs Portland Health Care System, 6Fondation LeDucq Transatlantic Networks of Excellence, 7Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medical Center The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures. Medicine Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury Kai-Zheong Lim*1, Christopher D. Daly*1,2,3, Peter Ghosh4, Graham Jenkin3, David Oehme5, Justin Cooper-White6,7, Taryn Naidoo6, Tony Goldschlager1,8 1Department of Surgery, Monash University, 2Department of Neurosurgery, Monash University, 3The Ritchie Centre, Hudson Institute of Medical Research, 4Proteobioactives, Pty Ltd, 5 Intervertebral disc degeneration is a significant contributor to back pain and a leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration exist. We demonstrate an ovine model of intervertebral disc degeneration, utilizing a drill bit, which achieves a consistent disc injury and reproducible level of disc degeneration.