Abstract

Regeneration of the intestinal epithelium after injury or during pathogenesis is a dynamic cellular process critical for host immunity. However, current epithelial injury models provide poor spatial control, complicating the study of precise cellular responses. Here we developed endoscopic femtosecond-laser surgery capable of generating acute tissue injury. A side-view probe provides a convenient access to the distal colon in the mouse in vivo and allows real-time intraoperative monitoring as well as pre- and post-surgery examinations via multiphoton imaging. The photo-induced damage showed a nonlinear dependence on laser intensity. At an optical power of 200 mW (2.5 nJ per pulse), scanning the beam focus over 300x300 µm2 area in the colonic mucosa generated substantial vascular damages within 30 s. We confirmed the localized tissue damage and the physiologic regeneration of the disrupted epithelium by in situ barrier function assays, validating the animal model for epithelial regeneration following injury. The femtosecond endosurgery technique is applicable to various experimental models based on laser-induced perturbations.

© 2013 Optical Society of America

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  1. H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).
  2. M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
    [CrossRef] [PubMed]
  3. T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
    [CrossRef] [PubMed]
  4. H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
    [CrossRef] [PubMed]
  5. L. Li and H. Clevers, “Coexistence of quiescent and active adult stem cells in mammals,” Science327(5965), 542–545 (2010).
    [CrossRef] [PubMed]
  6. S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
    [CrossRef] [PubMed]
  7. W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
    [CrossRef] [PubMed]
  8. H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
    [CrossRef] [PubMed]
  9. W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
    [CrossRef] [PubMed]
  10. M. Antkowiak, M. L. Torres-Mapa, K. Dholakia, and F. J. Gunn-Moore, “Quantitative phase study of the dynamic cellular response in femtosecond laser photoporation,” Biomed. Opt. Express1(2), 414–424 (2010).
    [CrossRef] [PubMed]
  11. W. Watanabe, N. Arakawa, S. Matsunaga, T. Higashi, K. Fukui, K. Isobe, and K. Itoh, “Femtosecond laser disruption of subcellular organelles in a living cell,” Opt. Express12(18), 4203–4213 (2004).
    [CrossRef] [PubMed]
  12. N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
    [CrossRef] [PubMed]
  13. M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
    [CrossRef] [PubMed]
  14. M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
    [CrossRef] [PubMed]
  15. M. Q. Salomão and S. E. Wilson, “Femtosecond laser in laser in situ keratomileusis,” J. Cataract Refract. Surg.36(6), 1024–1032 (2010).
    [CrossRef] [PubMed]
  16. J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
    [CrossRef] [PubMed]
  17. V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
    [CrossRef] [PubMed]
  18. P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
    [CrossRef] [PubMed]
  19. A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
    [CrossRef]
  20. C. B. Schaffer, A. Brodeur, J. F. García, and E. Mazur, “Micromachining bulk glass by use of femtosecond laser pulses with nanojoule energy,” Opt. Lett.26(2), 93–95 (2001).
    [CrossRef] [PubMed]
  21. A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
    [CrossRef] [PubMed]

2013 (2)

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

2012 (2)

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

2011 (1)

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

2010 (5)

M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
[CrossRef] [PubMed]

M. Q. Salomão and S. E. Wilson, “Femtosecond laser in laser in situ keratomileusis,” J. Cataract Refract. Surg.36(6), 1024–1032 (2010).
[CrossRef] [PubMed]

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

L. Li and H. Clevers, “Coexistence of quiescent and active adult stem cells in mammals,” Science327(5965), 542–545 (2010).
[CrossRef] [PubMed]

M. Antkowiak, M. L. Torres-Mapa, K. Dholakia, and F. J. Gunn-Moore, “Quantitative phase study of the dynamic cellular response in femtosecond laser photoporation,” Biomed. Opt. Express1(2), 414–424 (2010).
[CrossRef] [PubMed]

2009 (3)

M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
[CrossRef] [PubMed]

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

2008 (1)

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

2007 (1)

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

2006 (1)

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

2005 (1)

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

2004 (1)

2003 (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

2002 (1)

W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
[CrossRef] [PubMed]

2001 (1)

Abo, A.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Ajima, R.

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

Antkowiak, M.

Arakawa, N.

Barker, N.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Blinder, P.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

Blumberg, R. S.

W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
[CrossRef] [PubMed]

Brodeur, A.

Brown, S. L.

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Cao, Z.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Caterina, M. J.

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

Choi, C.

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
[CrossRef] [PubMed]

Choi, M.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
[CrossRef] [PubMed]

Chong, K.

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

Chung, E.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Clevers, H.

L. Li and H. Clevers, “Coexistence of quiescent and active adult stem cells in mammals,” Science327(5965), 542–545 (2010).
[CrossRef] [PubMed]

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Colonna, M.

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Conway, K. L.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Dholakia, K.

Friedman, B.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

Fukui, K.

Fukumura, D.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Fuss, I. J.

W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
[CrossRef] [PubMed]

García, J. F.

Geske, M. J.

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Gunn-Moore, F. J.

Higashi, T.

Hung, K. E.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Hüttman, G.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

Isobe, K.

Itoh, K.

Jain, R. K.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Jung, K.

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

Kim, J. K.

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

Kim, P.

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Kim, S.

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

King, L. S.

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

Kleinfeld, D.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

Ku, T.

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

Kucherlapati, R.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Kujala, P.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Lee, W. M.

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

Li, C.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Li, L.

L. Li and H. Clevers, “Coexistence of quiescent and active adult stem cells in mammals,” Science327(5965), 542–545 (2010).
[CrossRef] [PubMed]

Luo, C. T.

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

Lyden, P. D.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

Matsunaga, S.

Mazur, E.

Miyoshi, H.

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Mizoguchi, A.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Morin, B.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Neufert, C.

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

Neurath, M. F.

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

Nishimura, N.

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

Noack, J.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

Paltauf, G.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

Patel, K. K.

M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
[CrossRef] [PubMed]

Peters, P. J.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Salomão, M. Q.

M. Q. Salomão and S. E. Wilson, “Femtosecond laser in laser in situ keratomileusis,” J. Cataract Refract. Surg.36(6), 1024–1032 (2010).
[CrossRef] [PubMed]

Sato, T.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Schaffer, C. B.

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

C. B. Schaffer, A. Brodeur, J. F. García, and E. Mazur, “Micromachining bulk glass by use of femtosecond laser pulses with nanojoule energy,” Opt. Lett.26(2), 93–95 (2001).
[CrossRef] [PubMed]

Schweitzer, K. S.

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

Seno, H.

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Shi, H. N.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Shih, A. Y.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

Shimoda, L.

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

Sidhaye, V. K.

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

Snippert, H. J.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Sokol, H.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Stange, D. E.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Stanley, G.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

Stappenbeck, T. S.

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
[CrossRef] [PubMed]

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Strober, W.

W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
[CrossRef] [PubMed]

Torres-Mapa, M. L.

Tsai, P. S.

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

van de Wetering, M.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

van Es, J. H.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Villablanca, E. J.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Vogel, A.

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

Vries, R. G.

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Walker, M. R.

M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
[CrossRef] [PubMed]

Watanabe, W.

Webb, W. W.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Weigmann, B.

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

Wijmenga, C.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Williams, R. M.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Wilson, S. E.

M. Q. Salomão and S. E. Wilson, “Femtosecond laser in laser in situ keratomileusis,” J. Cataract Refract. Surg.36(6), 1024–1032 (2010).
[CrossRef] [PubMed]

Wirtz, S.

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

Xavier, R. J.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Yamaguchi, T. P.

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

Yamashita, H.

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Yoon, J.

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
[CrossRef] [PubMed]

Yun, S. H.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Zhang, M.

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Annu. Rev. Immunol. (1)

W. Strober, I. J. Fuss, and R. S. Blumberg, “The immunology of mucosal models of inflammation,” Annu. Rev. Immunol.20(1), 495–549 (2002).
[CrossRef] [PubMed]

Appl. Phys. B (1)

A. Vogel, J. Noack, G. Hüttman, and G. Paltauf, “Mechanisms of femtosecond laser nanosurgery of cells and tissues,” Appl. Phys. B81(8), 1015–1047 (2005).
[CrossRef]

Biomed. Opt. Express (1)

Gastroenterology (1)

H. Sokol, K. L. Conway, M. Zhang, M. Choi, B. Morin, Z. Cao, E. J. Villablanca, C. Li, C. Wijmenga, S. H. Yun, H. N. Shi, and R. J. Xavier, “Card9 mediates intestinal epithelial cell restitution, T-helper 17 responses, and control of bacterial infection in mice,” Gastroenterology145, 591–601 (2013).

J. Biomed. Opt. (1)

M. Choi, J. Yoon, and C. Choi, “Label-free optical control of arterial contraction,” J. Biomed. Opt.15(1), 015006 (2010).
[CrossRef] [PubMed]

J. Cataract Refract. Surg. (1)

M. Q. Salomão and S. E. Wilson, “Femtosecond laser in laser in situ keratomileusis,” J. Cataract Refract. Surg.36(6), 1024–1032 (2010).
[CrossRef] [PubMed]

J. Pathol. (1)

M. R. Walker, K. K. Patel, and T. S. Stappenbeck, “The stem cell niche,” J. Pathol.217(2), 169–180 (2009).
[CrossRef] [PubMed]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol.21(11), 1369–1377 (2003).
[CrossRef] [PubMed]

Nat. Methods (2)

N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, “Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke,” Nat. Methods3(2), 99–108 (2006).
[CrossRef] [PubMed]

P. Kim, E. Chung, H. Yamashita, K. E. Hung, A. Mizoguchi, R. Kucherlapati, D. Fukumura, R. K. Jain, and S. H. Yun, “In vivo wide-area cellular imaging by side-view endomicroscopy,” Nat. Methods7(4), 303–305 (2010).
[CrossRef] [PubMed]

Nat. Neurosci. (1)

A. Y. Shih, P. Blinder, P. S. Tsai, B. Friedman, G. Stanley, P. D. Lyden, and D. Kleinfeld, “The smallest stroke: occlusion of one penetrating vessel leads to infarction and a cognitive deficit,” Nat. Neurosci.16(1), 55–63 (2013).
[CrossRef] [PubMed]

Nat. Protoc. (2)

S. Wirtz, C. Neufert, B. Weigmann, and M. F. Neurath, “Chemically induced mouse models of intestinal inflammation,” Nat. Protoc.2(3), 541–546 (2007).
[CrossRef] [PubMed]

J. K. Kim, W. M. Lee, P. Kim, M. Choi, K. Jung, S. Kim, and S. H. Yun, “Fabrication and operation of GRIN probes for in vivo fluorescence cellular imaging of internal organs in small animals,” Nat. Protoc.7(8), 1456–1469 (2012).
[CrossRef] [PubMed]

Nature (1)

T. Sato, R. G. Vries, H. J. Snippert, M. van de Wetering, N. Barker, D. E. Stange, J. H. van Es, A. Abo, P. Kujala, P. J. Peters, and H. Clevers, “Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche,” Nature459(7244), 262–265 (2009).
[CrossRef] [PubMed]

Opt. Express (1)

Opt. Lett. (1)

Proc. Natl. Acad. Sci. U. S. A. (3)

V. K. Sidhaye, K. S. Schweitzer, M. J. Caterina, L. Shimoda, and L. S. King, “Shear stress regulates aquaporin-5 and airway epithelial barrier function,” Proc. Natl. Acad. Sci. U. S. A.105(9), 3345–3350 (2008).
[CrossRef] [PubMed]

M. Choi, T. Ku, K. Chong, J. Yoon, and C. Choi, “Minimally invasive molecular delivery into the brain using optical modulation of vascular permeability,” Proc. Natl. Acad. Sci. U. S. A.108(22), 9256–9261 (2011).
[CrossRef] [PubMed]

H. Seno, H. Miyoshi, S. L. Brown, M. J. Geske, M. Colonna, and T. S. Stappenbeck, “Efficient colonic mucosal wound repair requires Trem2 signaling,” Proc. Natl. Acad. Sci. U. S. A.106(1), 256–261 (2009).
[CrossRef] [PubMed]

Science (2)

H. Miyoshi, R. Ajima, C. T. Luo, T. P. Yamaguchi, and T. S. Stappenbeck, “Wnt5a potentiates TGF-β signaling to promote colonic crypt regeneration after tissue injury,” Science338(6103), 108–113 (2012).
[CrossRef] [PubMed]

L. Li and H. Clevers, “Coexistence of quiescent and active adult stem cells in mammals,” Science327(5965), 542–545 (2010).
[CrossRef] [PubMed]

Supplementary Material (1)

» Media 1: MOV (5651 KB)     

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Figures (7)

Fig. 1
Fig. 1

(a) A schematic of the multiphoton imaging and endosurgery system. Two-photon fluorescence images are displayed in a monitor in real time at 30 frames per second. The typical laser power at the tissue was adjusted to be 20-40 mW (i.e. 0.25-0.5 nJ) for noninvasive imaging and increased to 200 mW (i.e. 2.5 nJ) for inducing tissue damage. Displayed in the monitor in this illustration is a fluorescence image of the typical blood vessels in the colonic mucosa of a mouse. Scale bar, 100 µm. (b) Illustration of femtosecond laser endosurgery in the colon by the side-view probe.

Fig. 2
Fig. 2

Laser-induced damage on the mucosal blood vessels. (a) Time-series vascular images during femtosecond endosurgery (Media 1, 5.5 MB). Note that formation of intravascular clots (white arrows) and leakage of the vascular dye (yellow arrows). Scale bar, 50 µm. (b) Quantification of the blood flow before and after the laser irradiation in (a).

Fig. 3
Fig. 3

Power dependence of the laser-induced tissue injury. (a) The percentage of non-functional vascular segments in the FOV (300 × 300 μm2) for 100 mW and 200 mW, measured at 5 sec interval (n = 75-92). (b) The percentage of non-functional vascular segments after exposure for 60 s (n = 8). Over 60 s, the effective irradiation time on each focal point (1 µm2) is just 0.67 ms. Line, a Sigmoidal curve fit (R2 = 0.97).

Fig. 4
Fig. 4

Laser-induced cell death. (a) Bright-field images of excised tissues. (b) Fluorescence images of the tissues. Dead cells are identified by red fluorescence from Ethidium homodimer. The white dotted rectangle corresponds to the elongated irradiation region achieved by translating the probe along the lumen. Scale bars, 500 µm.

Fig. 5
Fig. 5

Evaluation of the epithelial barrier function. (a) A photograph showing Evans blue in the laser-irradiated region (arrow). (b) Bright-field images of the intact (control) and laser-injured (irradiated) areas. (c) Fluorescence (fluorescein-conjugated dextran) images of the intact and irradiated areas. Scale bars, 5 mm in (a) and 500 µm in (b-c).

Fig. 6
Fig. 6

Longitudinal follow-up imaging in vivo. (a) Experimental design. (b) Images acquired from a mouse at pre-op, post-op, and 2 days after the laser injury. Scale bar, 50 µm.

Fig. 7
Fig. 7

Epithelial regeneration after laser-induced injury. (a) Bright-field images taken with ex vivo preparation at 0, 1, and 4 days after laser endosurgery. The penetration of Evans blue indicates the disruption of the epithelial barrier function. Scale bar, 500 µm. (b) The measured tissue area stained by Evans blue. Error bars, standard deviations (n = 3).

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