title: Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses.
abstract: Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent re-oxygenation of the tissues (reperfusion). In the skin, ischemia-reperfusion (IR) is the main contributing factor to the pathophysiology of pressure ulcers. While the cascade of events leading up to the inflammatory response has been well studied, the spatial and temporal responses of the different subsets of immune cells to an IR injury are not well understood. Existing models of IR using the clamping technique on the skin flank are highly invasive and unsuitable for studying immune responses to injury, while similar non-invasive magnet clamping studies in the skin flank are less-than-ideal for intravital imaging studies. In this protocol, we describe a robust model of non-invasive IR developed on mouse ear skin, where we aim to visualize in real-time the cellular response of immune cells after reperfusion via multiphoton intravital imaging (MP-IVM).
authors.affiliation: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis; Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore.
authors.forename: Chi Ching
authors.initials: CC
authors.lastname: Goh
authors.affiliation: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis.
authors.forename: Jackson LiangYao
authors.initials: JL
authors.lastname: Li
authors.affiliation: Lee Kong Chian School of Medicine, Nanyang Technological University.
authors.forename: David
authors.initials: D
authors.lastname: Becker
authors.affiliation: Centenary Institute for Cancer Medicine and Cell Biology; Discipline of Dermatology, University of Sydney; Department of Dermatology, Royal Prince Alfred Hospital.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Weninger
authors.affiliation: Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore; LSI Immunology Programme, National University of Singapore.
authors.forename: Veronique
authors.initials: V
authors.lastname: Angeli
authors.affiliation: Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis; Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore; Discipline of Dermatology, University of Sydney; School of Biological Sciences, Nanyang Technological University; Ng_Lai_Guan@immunol.a-star.edu.sg.
authors.forename: Lai Guan
authors.initials: LG
authors.lastname: Ng
mesh_terms: D000818:Animals; D003250:Constriction; D004423:Ear; D006439:Hemodynamics; D007111:Immunity, Cellular; D000069416:Intravital Microscopy; D051379:Mice; D015427:Reperfusion Injury / Q000000981:diagnostic imaging*; D012867:Skin / Q000276:immunology / Q000503:physiopathology*