Multiphoton Intravital Microscopy of the Transplanted Mouse Kidney

• Published in: American journal of transplantation Vol. 11; no. 10; pp. 2067 - 2074
• Main Authors: Camirand, G., Li, Q., Demetris, A. J., Watkins, S. C., Shlomchik, W. D., Rothstein, D. M., Lakkis, F. G.
• Format: Journal Article
• Language: English
• Published: Malden, USA Blackwell Publishing Inc 01.10.2011; Wiley
• Subjects: Animals; Biological and medical sciences; Intravital imaging; ischemia/reperfusion injury; Kidney - metabolism; Kidney - pathology; kidney graft function; Kidney Transplantation; Medical sciences; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence; mouse kidney transplantation; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Surgery of the urinary system; Vitreous Body; Graft(material); kidney graft function; Temperature; Renal function; Two photon microscopy; ischemia/reperfusion injury; Environmental factor; Cardiovascular disease; Homotransplantation; Kidney; Reperfusion; Ischemia; Surgery; Graft; Intravital imaging; Cold; Rodentia; mouse kidney transplantation; Vertebrata; Mammalia; Treatment; Urinary system; Mouse; Animal; Medical imagery; Kidney transplantation
• ISSN: 1600-6135; 1600-6143
• DOI: 10.1111/j.1600-6143.2011.03671.x

Graft outcomes after kidney transplantation continue to be adversely affected by ischemia‐reperfusion injury and rejection. High‐resolution, real‐time imaging of the transplanted kidney could shed valuable insights into these dynamic processes, but such methodology has not been established. Here we describe a technique for intravital imaging of the transplanted mouse kidney using multiphoton fluorescence microscopy. The technique enabled real‐time, high‐resolution imaging and quantitation of renal filtration, cell death, leukocyte adhesion and capillary blood flow after transplantation. Using this technique, we found that brief graft ischemia associated with the transplantation procedure led to a rapid decline in renal filtration accompanied by a significant increase in microvascular leakage and renal tubular epithelial cell death within the first 3 h after transplantation. No significant changes in leukocyte adhesion or capillary blood flow were observed during the same time period. This report establishes multiphoton fluorescence microscopy as a sensitive tool for simultaneously studying functional and structural perturbations that occur in the mouse kidney after transplantation and for investigating the migration of leukocytes to the graft. This article reports the methodology to perform multiphoton intravital microscopy imaging of transplanted kidneys in mice and measures the early effects of the transplantation procedure on the graft.