title: Three-dimensional reconstruction of extravascular matrix patterns and blood vessels in human uveal melanoma tissue: techniques and preliminary findings.
abstract: OBJECTIVE
  Looping patterns rich in laminin are present in tissue samples of primary aggressive human uveal melanomas and their metastases. Because these extravascular patterns connect to blood vessels and transmit fluid in vitro and in vivo, the three-dimensional configuration of these patterns has been the subject of considerable speculation. In the current study, methods were devised to describe the three-dimensional configuration of looping extravascular matrix patterns in archival human uveal melanoma tissue.
  
  
  METHODS
  Twenty-five serial 4-microm-thick sections from primary uveal melanoma tissue were labeled with fluorescence-tagged laminin and examined by confocal microscopy to generate a Z-series within each 4-microm-thick section. The z-series from each section was stacked using an immersive three-dimensional environment (ImmersaDesk; Fakespace, Kitchener, Ontario, Canada) to allow for precise alignment and compensation for distortion artifact.
  
  
  RESULTS
  Extravascular matrix patterns that appeared to form loops in two dimensions were shown to represent thin wrappings around branching and twisting cylindrical groupings of melanoma cells. Blood vessels joined with some of these laminin-positive cylindrical wrappings.
  
  
  CONCLUSIONS
  In this preliminary study, periodic acid-Schiff (PAS)-positive laminin-rich looping patterns in two-dimensional tissue sections appear to outline cylindrical branching packets of melanoma cells rather than spheroidal nests. The conduction of fluid through this extravascular system may provide a novel delivery system for contrast and diagnostic agents.
authors.affiliation: Department of Pathology, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
authors.forename: Xue
authors.initials: X
authors.lastname: Chen
authors.forename: Zhuming
authors.initials: Z
authors.lastname: Ai
authors.forename: Mary
authors.initials: M
authors.lastname: Rasmussen
authors.forename: Peter
authors.initials: P
authors.lastname: Bajcsy
authors.forename: Loretta
authors.initials: L
authors.lastname: Auvil
authors.forename: Michael
authors.initials: M
authors.lastname: Welge
authors.forename: Lu
authors.initials: L
authors.lastname: Leach
authors.forename: Sumalee
authors.initials: S
authors.lastname: Vangveeravong
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Maniotis
authors.forename: Robert
authors.initials: R
authors.lastname: Folberg
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods; D007797:Laminin / Q000378:metabolism; D008545:Melanoma / Q000098:blood supply*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009389:Neovascularization, Pathologic / Q000473:pathology*; D014604:Uveal Neoplasms / Q000098:blood supply*
chemical_list: D018952:Antigens, CD34; D007797:Laminin