title: The use of Masson's trichrome staining, second harmonic imaging and two-photon excited fluorescence of collagen in distinguishing intestinal tuberculosis from Crohn's disease.
abstract: OBJECTIVE
  Differentiation between Crohn's disease (CD) and intestinal tuberculosis (ITB) continues to be difficult. The present study investigated the collagen fibre characteristics of CD and ITB using Masson's trichrome staining, second harmonic generation (SHG) imaging and two-photon excited fluorescence (TPEF) imaging with the aim of distinguishing between them.
  
  
  METHODS
  The characteristics of collagen fibres in intestinal specimens from patients with CD, ITB and healthy controls were compared using Masson's trichrome staining and SHG and TPEF imaging.
  
  
  RESULTS
  Masson's trichrome staining showed that the content of collagen fibre (540.92 [139.61-1681.93] vs 236.17 [72.94-1108.32], P < 0.05) and fibre deposits (888.92 [315.89-3172.9] vs 498.98 [38.82-5802.31], P < 0.05) were both higher in ITB than in CD. The content of collagen fibre (594.677 [139.61-1681.93] vs 107.425 [4.66-988.7], P < 0.05) and fibre deposits (1118.4661 [315.89-5802.31] vs 340.575 [29.62-1188.87], P < 0.05) were significantly higher in lesions with granulomata than in those without. The SHG/TPEF images demonstrated that the percentage of fibrosis in ITB was also significantly higher than in CD (P < 0.05), in both surgical (13.363% ± 5.303% vs 8.322% ± 5.078%, P = 0.044) and endoscopic specimens (mean rank 13.5 vs 7.5, P = 0.023). The SHG/TPEF imaging described different distribution patterns of collagen between CD and ITB; in the former this was irregular in clumps while in ITB the collagen was arranged around caseating granulomata.
  
  
  CONCLUSIONS
  The evaluation of fibrosis in CD and ITB by Masson's trichrome staining and SHG and TPEF imaging appears to distinguish between these two diseases.
authors.affiliation: Department of Digestive Disease, Zhujiang Hosiptal of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: H
authors.initials: H
authors.lastname: Mao
authors.affiliation: Department of Digestive Disease, Zhujiang Hosiptal of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: P
authors.initials: P
authors.lastname: Su
authors.affiliation: Department of Digestive Disease, Guangzhou Hospital of Traditional Chinese and Western Medicine, Guangzhou, Guangdong, China.
authors.forename: W
authors.initials: W
authors.lastname: Qiu
authors.affiliation: Department of Digestive Disease, Zhujiang Hosiptal of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: L
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Digestive Disease, Zhujiang Hosiptal of Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: H
authors.initials: H
authors.lastname: Yu
authors.affiliation: Research Centre of Non-labeled Imaging, Southern Medical University, Guangzhou, Guangdong, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
mesh_terms: D001391:Azo Compounds; D016022:Case-Control Studies; D003094:Collagen / Q000378:metabolism; D003424:Crohn Disease / Q000175:diagnosis*; D003937:Diagnosis, Differential; D004801:Eosine Yellowish-(YS); D006801:Humans; D007410:Intestinal Diseases / Q000175:diagnosis* / Q000382:microbiology; D008739:Methyl Green; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*; D014385:Tuberculosis, Gastrointestinal / Q000175:diagnosis*
chemical_list: D001391:Azo Compounds; C017768:trichrome stain; D008739:Methyl Green; D003094:Collagen; D004801:Eosine Yellowish-(YS)
keywords: Crohn's disease; collagen; intestinal tuberculosis; second harmonic generation