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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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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

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Conflict of Interest Statement

None declared.

Author information

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 (2011). https://doi.org/10.1007/s11605-011-1632-5

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Keywords

  • Two-photon laser scanning microscopy
  • Green fluorescent protein
  • Dextran sulfate sodium
  • Colitis
  • Corticosteroid