title: 3-dimensional digital reconstruction of the murine coronary system for the evaluation of chronic allograft vasculopathy.
abstract: BACKGROUND
  Chronic allograft vasculopathy (CAV) is a major mechanism of graft failure of transplanted organs in humans. Morphometric analysis of coronary arteries enables the quantitation of CAV in mouse models of heart transplantation. However, conventional histological procedures using single 2-dimensional sections limit the accuracy of CAV quantification. The aim of this study is to improve the accuracy of CAV quantification by reconstructing the murine coronary system in 3-dimensions (3D) and using virtual reconstruction and volumetric analysis to precisely assess neointimal thickness.
  
  
  METHODS
  Mouse tissue samples, native heart and transplanted hearts with chronic allograft vasculopathy, were collected and analyzed. Paraffin embedded samples were serially sectioned, stained and digitized using whole slide digital imaging techniques under normal and ultraviolet lighting. Sophisticated software tools were used to generate and manipulate 3D reconstructions of the major coronary arteries and branches.
  
  
  RESULTS
  The 3D reconstruction provides not only accurate measurements but also exact volumetric data of vascular lesions. This virtual coronary arteriography demonstrates that the vasculopathy lesions in this model are localized to the proximal coronary segments. In addition, virtual rotation and volumetric analysis enabled more precise measurements of CAV than single, randomly oriented histologic sections, and offer an improved readout for this important experimental model.
  
  
  CONCLUSIONS
  We believe 3D reconstruction of 2D histological slides will provide new insights into pathological mechanisms in which structural abnormalities play a role in the development of a disease. The techniques we describe are applicable to the analysis of arteries, veins, bronchioles and similar sized structures in a variety of tissue types and disease model systems.
  The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/3772457541477230 .
authors.affiliation: 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary. fonyadla@gmail.com.
authors.forename: László
authors.initials: L
authors.lastname: Fónyad
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. kshino49@yahoo.co.jp.
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Shinoda
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. efarkash@med.umich.edu.
authors.forename: Evan A
authors.initials: EA
authors.lastname: Farkash
authors.affiliation: microDimensions GmbH, Munich, Germany. martin.groher@micro-dimensions.com.
authors.forename: Martin
authors.initials: M
authors.lastname: Groher
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. DSEBASTIAN@partners.org.
authors.forename: Divya P
authors.initials: DP
authors.lastname: Sebastian
authors.affiliation: 2nd Department of Pathology, Semmelweis University, Budapest, Hungary. cac@korb2.sote.hu.
authors.forename: A Marcell
authors.initials: AM
authors.lastname: Szász
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. COLVIN@helix.mgh.harvard.edu.
authors.forename: Robert B
authors.initials: RB
authors.lastname: Colvin
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. YYAGI@partners.org.
authors.forename: Yukako
authors.initials: Y
authors.lastname: Yagi
mesh_terms: D000465:Algorithms; D064591:Allografts; D015901:Angiography, Digital Subtraction; D000818:Animals; D002908:Chronic Disease; D017023:Coronary Angiography; D003324:Coronary Artery Disease / Q000209:etiology / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D003331:Coronary Vessels / Q000378:metabolism / Q000473:pathology*; D004195:Disease Models, Animal; D005240:Feasibility Studies; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D016027:Heart Transplantation / Q000009:adverse effects*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D058426:Neointima; D016612:Paraffin Embedding; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D012984:Software
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins