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University College Cork

8 ARTICLES PUBLISHED IN JoVE

Medicine

Murine Bioluminescent Hepatic Tumour Model

Simon Rajendran¹, Slawomir Salwa¹, Xuefeng Gao², Sabin Tabirca², Deirdre O'Hanlon³, Gerald C. O'Sullivan¹, Mark Tangney¹

¹Cork Cancer Research Centre, Mercy University Hospital and Leslie C. Quick Jnr. Laboratory, University College Cork, ²Department of Computer Science, University College Cork, ³South Infirmary Victoria University Hospital

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Medicine

Ex Vivo Culture of Patient Tissue & Examination of Gene Delivery

Simon Rajendran¹, Slawomir Salwa¹, Xuefeng Gao², Sabin Tabirca², Deirdre O'Hanlon², Gerald C. O'Sullivan¹, Mark Tangney¹

¹Cork Cancer Research Centre, Mercy University Hospital and Leslie C. Quick Jnr. Laboratory, University College Cork, ²Department of Computer Science, University College Cork

This article describes the culture of patient tissue slices for gene delivery studies and subsequent analysis of gene expression using IVIS bioluminescence imaging.

Immunology and Infection

Bioluminescent Bacterial Imaging In Vivo

Chwanrow K. Baban^*1, Michelle Cronin^*1, Ali R. Akin¹, Anne O'Brien¹, Xuefeng Gao¹, Sabin Tabirca¹, Kevin P. Francis¹, Mark Tangney¹

¹Cork Cancer Research Centre, BioSciences Institute, University College Cork

This article describes the administration of lux-tagged bacteria to mice and subsequent in vivo analysis using IVIS bioluminescence imaging.

Medicine

Combined In vivo Optical and µCT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Nicholas M. Bernthal¹, Brad N. Taylor², Jeffrey A. Meganck², Yu Wang³, Jonathan H. Shahbazian³, Jared A. Niska¹, Kevin P. Francis², Lloyd S. Miller^3,4

¹Orthopaedic Hospital Research Center, Orthopaedic Hospital Department of Orthopaedic Surgery, David Geffen School of Medicine at University of California, Los Angeles (UCLA), ²PerkinElmer, ³Department of Dermatology, Johns Hopkins University School of Medicine, ⁴Department of Medicine, Division of Infectious Diseases, Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine

Combined optical and μCT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

Immunology and Infection

Development of an Antigen-driven Colitis Model to Study Presentation of Antigens by Antigen Presenting Cells to T Cells

Valerio Rossini¹, Katarina Radulovic², Christian U. Riedel³, Jan Hendrik Niess²

¹APC Microbiome Institute, University College Cork, ²Division of Gastroenterology and Hepatology, University Hospital Basel, ³Institute of Microbiology and Biotechnology, University of Ulm

In this antigen-driven colitis model, OT-II CD4⁺ T cells expressing a red fluorescent protein were adoptively transferred into RAG^-/- mice that express a green fluorescent protein in mononuclear phagocytes (MPs). The hosts were challenged with Escherichia coli (E.coli) expressing the ovalbumin protein (OVA) fused to a cyan fluorescent protein (CFP).

Cancer Research

Utilizing ¹⁸F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer

Milica Momcilovic¹, Sean T. Bailey², Jason T. Lee³, Charles Zamilpa³, Anthony Jones³, Gihad Abdelhady¹, James Mansfield⁴, Kevin P. Francis⁵, David B. Shackelford¹

¹Division of Pulmonary and Critical Care Medicine, University of California Los Angeles David Geffen School of Medicine, ²University of North Carolina at Chapel Hill, ³Department of Molecular and Medical Pharmacology, University of California Los Angeles, ⁴Andor Technology, ⁵Division of Orthopaedic Surgery, University of California Los Angeles David Geffen School of Medicine

In this protocol, we describe how to utilize [¹⁸F]-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography (¹⁸F-FDG PET/CT) imaging to measure the tumor metabolic response to the targeted therapy MLN0128 in a Kras/Lkb1 mutant mouse model of lung cancer and coupled imaging with high resolution ex vivo autoradiography and quantitative histology.

Biology

Fluorescence Lifetime Macro Imager for Biomedical Applications

Rajannya Sen¹, Alexander V. Zhdanov¹, Ciaran Devoy², Mark Tangney², Liisa M. Hirvonen³, Andrei Nomerotski⁴, Dmitri B. Papkovsky¹

¹School of Biochemistry and Cell Biology, University College Cork, ²Cancer Research@UCC, University College Cork, ³Centre for Microscopy, Characterisation and Analysis (CMCA), The University of Western Australia, ⁴Physics Department, Brookhaven National Laboratory

This paper describes the use of a new, fast optical imager for the macroscopic photoluminescence lifetime imaging of long decay emitting samples. The integration, image acquisition, and analysis procedures are described, along with the preparation and characterization of the sensor materials for the imaging and the application of the imager in studying biological samples.

Biology

Quantification of Cytokine-Induced Cell Death in Human Colonic Organoids Using Live Fluorescence Microscopy

Peter Flood¹, Ken Nally^1,2

¹APC Microbiome Ireland, University College Cork, ²School of Biochemistry and Cell Biology, University College Cork

This protocol describes a simple and cost-effective method to investigate and quantify cell death in human colonic organoids in response to cytotoxic perturbagens such as cytokines. The approach employs a fluorescent cell death dye (SYTOX Green Nucleic Acid Stain), live fluorescence microscopy, and open-source image analysis software to quantify single-organoid responses to cytotoxic stimuli.