title: Long term intravital multiphoton microscopy imaging of immune cells in healthy and diseased liver using CXCR6.Gfp reporter mice.
abstract: Liver inflammation as a response to injury is a highly dynamic process involving the infiltration of distinct subtypes of leukocytes including monocytes, neutrophils, T cell subsets, B cells, natural killer (NK) and NKT cells. Intravital microscopy of the liver for monitoring immune cell migration is particularly challenging due to the high requirements regarding sample preparation and fixation, optical resolution and long-term animal survival. Yet, the dynamics of inflammatory processes as well as cellular interaction studies could provide critical information to better understand the initiation, progression and regression of inflammatory liver disease. Therefore, a highly sensitive and reliable method was established to study migration and cell-cell-interactions of different immune cells in mouse liver over long periods (about 6 hr) by intravital two-photon laser scanning microscopy (TPLSM) in combination with intensive care monitoring. The method provided includes a gentle preparation and stable fixation of the liver with minimal perturbation of the organ; long term intravital imaging using multicolor multiphoton microscopy with virtually no photobleaching or phototoxic effects over a time period of up to 6 hr, allowing tracking of specific leukocyte subsets; and stable imaging conditions due to extensive monitoring of mouse vital parameters and stabilization of circulation, temperature and gas exchange. To investigate lymphocyte migration upon liver inflammation CXCR6.gfp knock-in mice were subjected to intravital liver imaging under baseline conditions and after acute and chronic liver damage induced by intraperitoneal injection(s) of carbon tetrachloride (CCl4). CXCR6 is a chemokine receptor expressed on lymphocytes, mainly on Natural Killer T (NKT)-, Natural Killer (NK)- and subsets of T lymphocytes such as CD4 T cells but also mucosal associated invariant (MAIT) T cells1. Following the migratory pattern and positioning of CXCR6.gfp+ immune cells allowed a detailed insight into their altered behavior upon liver injury and therefore their potential involvement in disease progression.
authors: Department of Medicine III, RWTH University-Hospital Aachen; fheymann@ukaachen.de.
authors: Felix
authors: F
authors: Heymann
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Patricia M
authors: PM
authors: Niemietz
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Julia
authors: J
authors: Peusquens
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Can
authors: C
authors: Ergen
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Marlene
authors: M
authors: Kohlhepp
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Jana C
authors: JC
authors: Mossanen
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Carlo
authors: C
authors: Schneider
authors: IZKF Aachen Core Facility "Two-Photon Imaging", RWTH University-Hospital Aachen.
authors: Michael
authors: M
authors: Vogt
authors: Institute for Laboratory Animal Science & Experimental Surgery, RWTH Aachen University.
authors: Rene H
authors: RH
authors: Tolba
authors: Department of Medicine III, RWTH University-Hospital Aachen.
authors: Christian
authors: C
authors: Trautwein
authors: Institute for Pharmacology, RWTH University-Hospital Aachen.
authors: Christian
authors: C
authors: Martin
authors: Department of Medicine III, RWTH University-Hospital Aachen; frank.tacke@gmx.net.
authors: Frank
authors: F
authors: Tacke
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000473:pathology; D002450:Cell Communication / Q000276:immunology; D002465:Cell Movement / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D008099:Liver / Q000166:cytology* / Q000276:immunology*; D008107:Liver Diseases / Q000276:immunology* / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D055611:Natural Killer T-Cells / Q000166:cytology / Q000473:pathology; D054387:Receptors, CXCR / Q000737:chemistry / Q000235:genetics; D000075744:Receptors, CXCR6; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D016176:T-Lymphocyte Subsets / Q000276:immunology
chemical_list: C482906:Cxcr6 protein, mouse; D054387:Receptors, CXCR; D000075744:Receptors, CXCR6; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins