title: Intravital three-dimensional dynamic pathology of experimental colitis in living mice using two-photon laser scanning microscopy.
abstract: BACKGROUND
  Intravital three-dimensional (3D) visualization of treatment efficacy in experimental colitis in living mice using two-photon laser scanning microscopy (TPLSM) has not been described.
  
  
  METHODS
  Colitis was induced with dextran sulfate sodium (DSS) in green fluorescent protein (GFP) transgenic mice. The 3D tomographic image of DSS-induced colitis with or without prednisolone was obtained intravitally using TPLSM. A serosal-approaching method was developed, by which we could observe all layers of the cecum from serosa to luminal mucosa without opening and everting the cecum. The dynamic pathology and treatment efficacy were assessed in the same mouse on several occasions.
  
  
  RESULTS
  The time-lapse 3D tomographic movie of DSS-induced colitis was obtained in living mice at a magnification of greater than ×600, which demonstrated irregularity of crypts, disappearance of crypts, inflammatory cell infiltrates in the lamina propria, and abscess formation at the bottom of crypts. Intravital TPLSM in the same mice demonstrated fewer infiltrating leukocytes and crypt abscesses on day 14 in the steroid group compared with the nonsteroid group.
  
  
  CONCLUSIONS
  Intravital 3D tomographic visualization of experimental colitis using TPLSM in combination with the serosal-approaching method can provide dynamic pathology at a high magnification, which may be useful in evaluating treatment efficacy in the same living mice.
authors.affiliation: Department of Gastrointestinal and Pediatric Surgery, Mie University, Mie, Japan.
authors.forename: Yuhki
authors.initials: Y
authors.lastname: Morimoto
authors.forename: Koji
authors.initials: K
authors.lastname: Tanaka
authors.forename: Yuji
authors.initials: Y
authors.lastname: Toiyama
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Inoue
authors.forename: Toshimitsu
authors.initials: T
authors.lastname: Araki
authors.forename: Keiichi
authors.initials: K
authors.lastname: Uchida
authors.forename: Kazushi
authors.initials: K
authors.lastname: Kimura
authors.forename: Akira
authors.initials: A
authors.lastname: Mizoguchi
authors.forename: Masato
authors.initials: M
authors.lastname: Kusunoki
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000627:therapeutic use; D003092:Colitis / Q000139:chemically induced / Q000188:drug therapy / Q000473:pathology*; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D049452:Green Fluorescent Proteins; D021621:Imaging, Three-Dimensional; D049408:Luminescent Agents; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D011239:Prednisolone / Q000627:therapeutic use
chemical_list: D000893:Anti-Inflammatory Agents; D049408:Luminescent Agents; D049452:Green Fluorescent Proteins; D016264:Dextran Sulfate; D011239:Prednisolone