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Intravital Three-Dimensional Dynamic Pathology of Experimental Colitis in Living Mice Using Two-Photon Laser Scanning Microscopy

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Journal of Gastrointestinal Surgery

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.

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Abbreviations

TPLSM:

Two-photon laser scanning microscopy

GFP:

Green fluorescent protein

DSS:

Dextran sulfate sodium

References

  1. Wang BG, König K, Halbhuber KJ. Two-photon microscopy of deep intravital tissues and its merits in clinical research. J Microsc. 2010;238:1–20. Review.

    Article  PubMed  CAS  Google Scholar 

  2. Quentmeier S, Denicke S, Gericke KH. Two-color two-photon fluorescence laser scanning microscopy. J Fluoresc. 2009;19:1037–1043.

    Article  PubMed  CAS  Google Scholar 

  3. Hänninen P, Soukka J, Soini JT. Two-photon excitation fluorescence bioassays. Ann N Y Acad Sci. 2008;1130:320–326. Review.

    Article  PubMed  Google Scholar 

  4. Benninger RK, Hao M, Piston DW. Multi-photon excitation imaging of dynamic processes in living cells and tissues. Rev Physiol Biochem Pharmacol. 2008;160:71–92. Review.

    Article  PubMed  CAS  Google Scholar 

  5. Schenke-Layland K, Riemann I, Damour O, et al. Two-photon microscopes and in vivo multiphoton tomographs—powerful diagnostic tools for tissue engineering and drug delivery. Adv Drug Deliv Rev. 2006;58:878–896.

    Article  PubMed  CAS  Google Scholar 

  6. Starodub OT, Demitrack ES, Baumgartner HK, et al. Disruption of the Cox-1 gene slows repair of microscopic lesions in the mouse gastric epithelium. Am J Physiol Cell Physiol. 2008;294:C223–C232.

    Article  PubMed  CAS  Google Scholar 

  7. Toiyama Y, Mizoguchi A, Okugawa Y, et. al. Intravital imaging of DSS-induced cecal mucosal damage in GFP-transgenic mice using two-photon microscopy. J Gastroenterol. 2010;45:544–553.

    Article  PubMed  Google Scholar 

  8. Fritsch Fredin M, Vidal A, Utkovic H, et al. The application and relevance of ex vivo culture systems for assessment of IBD treatment in murine models of colitis. Pharmacol Res. 2008;58:222–231.

    Article  PubMed  CAS  Google Scholar 

  9. Islam MS, Murata T, Fujisawa M, et al. Anti-inflammatory effects of phytosteryl ferulates in colitis induced by dextran sulphate sodium in mice. Br J Pharmacol. 2008;154:812–824.

    Article  PubMed  CAS  Google Scholar 

  10. Okabe M, Ikawa M, Kominami K, et al. 'Green mice' as a source of ubiquitous green cells. FEBS Lett. 1997;407:313–319.

    Article  PubMed  CAS  Google Scholar 

  11. Li WB, Zuo XL, Li CQ, et al. Diagnostic value of confocal laser endomicroscopy for gastric superficial cancerous lesions. Gut. 2011;60:299–306.

    Article  PubMed  Google Scholar 

  12. Moussata D, Goetz M, Gloeckner A, et al. Confocal laser endomicroscopy is a new imaging modality for recognition of intramucosal bacteria in inflammatory bowel disease in vivo. Gut. 2011;60:26–33.

    Article  PubMed  Google Scholar 

  13. Tang S, Jung W, McCormick D, et al. Design and implementation of fiber-based multiphoton endoscopy with microelectromechanical systems scanning. J Biomed Opt. 2009;14:034005.

    Article  PubMed  Google Scholar 

  14. Murari K, Zhang Y, Li S, Chen Y, Li MJ, Li X. Compensation-free, all-fiber-optic, two-photon endomicroscopy at 1.55 μm. Opt Lett. 2011;36:1299–1301.

    Article  PubMed  Google Scholar 

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Conflict of Interest Statement

None declared.

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Authors and Affiliations

  1. Department of Gastrointestinal and Pediatric Surgery, Graduate School of Medicine, Mie University, 2-174 Edobashi, Tsu, Mie, 514-8507, Japan

    Yuhki Morimoto, Koji Tanaka, Yuji Toiyama, Yasuhiro Inoue, Toshimitsu Araki, Keiichi Uchida & Masato Kusunoki

  2. Department of Neural Regeneration and Cell Communication, Graduate School of Medicine, Mie University, 2-174 Edobashi, Tsu, Mie, 514-8507, Japan

    Kazushi Kimura & Akira Mizoguchi

Authors
  1. Yuhki Morimoto
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  2. Koji Tanaka
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  3. Yuji Toiyama
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  4. Yasuhiro Inoue
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  5. Toshimitsu Araki
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  6. Keiichi Uchida
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  7. Kazushi Kimura
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  8. Akira Mizoguchi
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  9. Masato Kusunoki
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Corresponding author

Correspondence to Koji Tanaka.

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Morimoto, Y., Tanaka, K., Toiyama, Y. et al. Intravital Three-Dimensional Dynamic Pathology of Experimental Colitis in Living Mice Using Two-Photon Laser Scanning Microscopy. J Gastrointest Surg 15, 1842–1850 (2011). https://doi.org/10.1007/s11605-011-1632-5

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  • DOI: https://doi.org/10.1007/s11605-011-1632-5

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