title: Assessing large-vessel endothelial permeability using near-infrared fluorescence imaging--brief report.
abstract: OBJECTIVE
  Loss of endothelial barrier function in arterial blood vessels is characteristic of vascular pathologies, including atherosclerosis. Here, we present a near-infrared fluorescence (NIRF) imaging methodology for quantifying endothelial permeability and macromolecular uptake in large arteries in the mouse and evaluate its applicability for studying mechanisms of vascular inflammation.
  
  
  RESULTS
  To validate the NIRF methodology, macrovascular inflammation was induced in C57bl/6 mice by local tumor necrosis factor-α stimulation of the carotid artery or in apolipoprotein E-deficient mice by Western diet for 4 weeks. Evans blue dye, serving as plasma protein marker and fluorescent in the near-infrared spectrum, was given intravenously at different doses. Carotids and aorta were excised, and Evans blue dye fluorescence was assessed through whole vessel scan in an infrared imaging system. NIRF correlated to extraction-absorbance methodology for Evans blue dye quantification and was superior at discriminating plasma protein accumulation in tumor necrosis factor-α-stimulated carotids. NIRF allowed for focal quantification of increased arterial wall Evans blue dye uptake in (apolipoprotein E-deficient) mice. Importantly, NIRF left vessels intact for subsequent histological analysis or quantification of leukocyte subpopulations by flow cytometry.
  
  
  CONCLUSIONS
  The described NIRF methodology provides a sensitive and rapid tool to locate and quantify macromolecular uptake in the wall of arterial blood vessels in vascular pathologies in mice.
authors: From the Department of Physiology and Pharmacology (E.L., A.M.V.D.D., E.K., L.L.) and Department of Molecular Medicine and Surgery, Center of Molecular Medicine (E.K.), Karolinska Institutet, Stockholm, Sweden; Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany (O.S.); Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (O.S.); and German Centre for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany (O.S.). erik.lundeberg@ki.se.
authors: Erik
authors: E
authors: Lundeberg
authors: From the Department of Physiology and Pharmacology (E.L., A.M.V.D.D., E.K., L.L.) and Department of Molecular Medicine and Surgery, Center of Molecular Medicine (E.K.), Karolinska Institutet, Stockholm, Sweden; Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany (O.S.); Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (O.S.); and German Centre for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany (O.S.).
authors: Anne M
authors: AM
authors: Van Der Does
authors: From the Department of Physiology and Pharmacology (E.L., A.M.V.D.D., E.K., L.L.) and Department of Molecular Medicine and Surgery, Center of Molecular Medicine (E.K.), Karolinska Institutet, Stockholm, Sweden; Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany (O.S.); Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (O.S.); and German Centre for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany (O.S.).
authors: Ellinor
authors: E
authors: Kenne
authors: From the Department of Physiology and Pharmacology (E.L., A.M.V.D.D., E.K., L.L.) and Department of Molecular Medicine and Surgery, Center of Molecular Medicine (E.K.), Karolinska Institutet, Stockholm, Sweden; Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany (O.S.); Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (O.S.); and German Centre for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany (O.S.).
authors: Oliver
authors: O
authors: Soehnlein
authors: From the Department of Physiology and Pharmacology (E.L., A.M.V.D.D., E.K., L.L.) and Department of Molecular Medicine and Surgery, Center of Molecular Medicine (E.K.), Karolinska Institutet, Stockholm, Sweden; Institute for Cardiovascular Prevention, Ludwig-Maximilians University, Munich, Germany (O.S.); Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (O.S.); and German Centre for Cardiovascular Research (DZHK), Partner Site Munich Heart Alliance, Munich, Germany (O.S.).
authors: Lennart
authors: L
authors: Lindbom
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism*; D002199:Capillary Permeability; D002339:Carotid Arteries / Q000378:metabolism*; D004396:Coloring Agents / Q000008:administration & dosage / Q000378:metabolism; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism*; D005070:Evans Blue / Q000008:administration & dosage / Q000378:metabolism; D005260:Female; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D007275:Injections, Intravenous; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D061848:Optical Imaging / Q000379:methods*; D015203:Reproducibility of Results; D019265:Spectroscopy, Near-Infrared; D014409:Tumor Necrosis Factor-alpha
chemical_list: D001057:Apolipoproteins E; D004396:Coloring Agents; D014409:Tumor Necrosis Factor-alpha; D005070:Evans Blue
keywords: atherosclerosis; endothelium; imaging; inflammation; permeability