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Chinese Optics Letters

Chinese Optics Letters


  • Vol. 11, Iss. 1 — Jan. 1, 2013
  • pp: 011703–

Recent advances in two-photon imaging: technology developments and biomedical applications

Yu Chen, Hengchang Guo, Wei Gong, Luye Qin, Hossein Aleyasin, Rajiv R Ratan, Sunghee Cho, Jianxin Chen, and Shusen Xie  »View Author Affiliations

Chinese Optics Letters, Vol. 11, Issue 1, pp. 011703- (2013)

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In the past two decades, two-photon microscopy (TPM) transforms biomedical research, allowing nondestructive high-resolution fluorescent molecular imaging and label-free imaging in vivo and in real time. Here we review the recent advances of TPM technology including novel laser sources, new image acquisition paradigms, and microendoscopic imaging systems. Then, we survey the capabilities of TPM imaging of biological relevant molecules such as nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD), and reactive oxygen species (ROS). Biomedical applications of TPM in neuroscience and cancer detection are demonstrated.

© 2013 Chinese Optics Letters

OCIS Codes
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.2150) Medical optics and biotechnology : Endoscopic imaging
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.6510) Medical optics and biotechnology : Spectroscopy, tissue diagnostics
(180.0180) Microscopy : Microscopy
(190.0190) Nonlinear optics : Nonlinear optics
(190.4180) Nonlinear optics : Multiphoton processes
(180.4315) Microscopy : Nonlinear microscopy

Yu Chen, Hengchang Guo, Wei Gong, Luye Qin, Hossein Aleyasin, Rajiv R Ratan, Sunghee Cho, Jianxin Chen, and Shusen Xie, "Recent advances in two-photon imaging: technology developments and biomedical applications," Chin. Opt. Lett. 11, 011703- (2013)

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  1. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
  2. D. Kobat, M. E. Durst, N. Nishimura, A. W. Wong, C. B. Schaffer, and C. Xu, Opt. Express 17, 13354 (2009).
  3. D. Kobat, N. G. Horton, and C. Xu, J. Biomed. Opt. 16, 106014 (2011).
  4. Y. Chen, C. P. Liang, Y. Liu, A. H. Fischer, A. V. Parwani, and L. Pantanowitz, J. Pathol. Inform. 3, 22 (2012).
  5. W. R. Zipfel, R. M. Williams, and W. W. Webb, Nat. Biotechnol. 21, 1369 (2003).
  6. K. Svoboda and R. Yasuda, Neuron 50, 823 (2006).
  7. A. C. Kwan and Y. Dan, Current Biology 22, 1459 (2012).
  8. H. Guo, H. Aleyasin, S. Howard, B. C. Dickinson, R. Haskew-Layton, D. Kobat, V. S. Lin, D. Rivera, C. J. Chang, R. R. Ratan, and C. Xu, in Proceedings of Optical Molecular Probes, Imaging and Drug Delivery 2011 OtuD3 (2011).
  9. Y. Li, H. Lu, P. L. Cheng, S. Ge, H. Xu, S. H. Shi, and Y. Dan, Nature 486, 118 (2012).
  10. D. Huber, D. A. Gutnisky, S. Peron, D. H. O'Connor, J. S. Wiegert, L. Tian, T. G. Oertner, L. L. Looger, and K. Svoboda, Nature 484, 473 (2012).
  11. T. Ladewig, P. Kloppenburg, P. M. Lalley, W. R. Zipfel, W. W. Webb, and B. U. Keller, J. Physiol. 547, 775 (2003).
  12. M. E. McLellan, S. T. Kajdasz, B. T. Hyman, and B. J. Bacskai, J. Neurosci. 23, 2212 (2003).
  13. M. Meyer-Luehmann, T. L. Spires-Jones, C. Prada, M. Garcia-Alloza, A. de Calignon, A. Rozkalne, J. Koenigsknecht-Talboo, D. M. Holtzman, B. J. Bacskai, and B. T. Hyman, Nature 451, 720 (2008).
  14. M. C. Skala, K. M. Riching, A. Gendron-Fitzpatrick, J. Eickhoff, K. W. Eliceiri, J. G. White, and N. Ramanujam, Proc. Natl. Acad. Sci. USA 104, 19494 (2007).
  15. H.-C. Guo, K. Takashima, A. Sato, H. Yokoyama, M. Mure, Y. Iseki, and H. Tsubokawa, Proc. SPIE 6860, 686020 (2008).
  16. D. M. McDonald and P. L. Choyke, Nat. Med. 9, 713 (2003).
  17. W.Wang, J. B.Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Huttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall, and J. S. Condeelis, Cancer Res. 62, 6278 (2002).
  18. M. C. Skala, J. M. Squirrell, K. M. Vrotsos, V. C. Eickhoff, A. Gendron-Fitzpatrick, K. W. Eliceiri, and N. Ramanujam, Cancer Res. 65, 1180 (2005).
  19. R. K. Jain, L. L. Munn, and D. Fukumura, Nat. Rev. Cancer 2, 266 (2002).
  20. S. W. Perry, R. M. Burke, and E. B. Brown, Ann. Biomed. Eng. 40, 277 (2012).
  21. J. M. Squirrell, D. L. Wokosin, J. G. White, and B. D. Bavister, Nat. Biotechnol. 17, 763 (1999).
  22. T. H. Harris, E. J. Banigan, D. A. Christian, C. Konradt, E. D. TaitWojno, K. Norose, E. H. Wilson, B. John,W. Weninger, A. D. Luster, A. J. Liu, and C. A. Hunter, Nature 486, 545 (2012).
  23. H. D. Moreau, F. Lemaitre, E. Terriac, G. Azar, M. Piel, A.-M. Lennon-Dumenil, and P. Bousso, Immunity 37, 351 (2012).
  24. P. Steven, F. Bock, G. Huttmann, and C. Cursiefen, PLoS One 6, e26253 (2011).
  25. G. D. Victora, T. A. Schwickert, D. R. Fooksman, A. O. Kamphorst, M. Meyer-Hermann, M. L. Dustin, and M. C. Nussenzweig, Cell 143, 592 (2010).
  26. M. D. Duncan, J. Reintjes, and T. J. Manuccia, Opt. Lett. 7, 350 (1982).
  27. H. Yokoyama, A. Sato, H. C. Guo, K. Sato, M. Murc, and H. Tsubokawa, Opt. Express 16, 17752 (2008).
  28. S. Sakadzic, E. Roussakis, M. A. Yaseen, E. T. Mandeville, V. J. Srinivasan, K. Arai, S. Ruvinskaya, A. Devor, E. H. Lo, S. A. Vinogradov, and D. A. Boas, Nat. Methods 7, 755 (2010).
  29. X. K. Shu, A. Royant, M. Z. Lin, T. A. Aguilera, V. Lev-Ram, P. A. Steinbach, and R. Y. Tsien, Science 324, 804 (2009).
  30. N. C. Shaner, P. A. Steinbach, and R. Y. Tsien, Nat. Methods 2, 905 (2005).
  31. D. Shcherbo, I. I. Shemiakina, A. V. Ryabova, K. E. Luker, B. T. Schmidt, E. A. Souslova, T. V. Gorodnicheva, L. Strukova, K. M. Shidlovskiy, O. V. Britanova, A. G. Zaraisky, K. A. Lukyanov, V. B. Loschenov, G. D. Luker, and D. M. Chudakov, Nat. Methods 7, 827 (2010).
  32. P. S Weiss, ACS Nano 2, 1977 (2008).
  33. A. Roda, Anal. Bioanal. Chem. 396, 1619 (2010).
  34. D. R. Larson, W. R. Zipfel, R. M. Williams, S. W. Clark, M. P. Bruchez, F. W. Wise, and W. W. Webb, Science 300, 1434 (2003).
  35. J. Park, A. Estrada, K. Sharp, K. Sang, J. A. Schwartz, D. K. Smith, C. Coleman, J. D. Payne, B. A. Korgel, A. K. Dunn, and J. W. Tunnell, Opt. Express 16, 1590 (2008).
  36. L. Cao, X.Wang, M. J. Meziani, F. S. Lu, H. F. Wang, P. J. G. Luo, Y. Lin, B. A. Harruff, L. M. Veca, D. Murray, S. Y. Xie, and Y. P. Sun, J. Am. Chem. Soc. 129, 11318 (2007).
  37. A. H. Fu, W. W. Gu, C. Larabell, and A. P. Alivisatos, Curr. Opin. Neurobiol. 15, 568 (2005).
  38. R. E. Rumbaut and A. J. Sial, Microcirculation 6, 205 (1999).
  39. A. M. Derfus, W. C. W. Chan, and S. N. Bhatia, Nano Lett. 4, 11 (2004).
  40. M. P. Jain, A. O. Choi, K. D. Nebibert, and D.Maysinger, Nanomed. 4, 277 (2009).
  41. W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, Proc. Natl. Acad. Sci USA 100, 7075 (2003).
  42. G. C. Su, Y. H. Wei, and H. W. Wang, Opt. Express 19, 21145 (2011).
  43. H. W. Guo, C. T. Chen, Y. H. Wei, O. K. Lee, V. Gukassyan, F. J. Kao, and H. W. Wang, J. Biomed. Opt. 13, 050505 (2008).
  44. H. W. Guo, Y. H. Wei, and H. W. Wang, J. Biomed. Opt. 16, 068001 (2011).
  45. S. M. Zhuo, J. Yan, G. Chen, J. X. Chen, Y. C. Liu, J. P. Lu, X. Q. Zhu, X. S. Jiang, and S. S. Xie, Biomed. Opt. Express 2, 615 (2011).
  46. P. J. Campagnola and L. M. Loew, Nat. Biotechnol. 21, 1356 (2003).
  47. A. Zumbusch, G. R. Holtom, and X. S. Xie, Phys. Rev. Lett. 82, 4142 (1999).
  48. J.-X. Cheng and X. S. Xie, J. Phys. Chem. B 108, 827 (2004).
  49. C. Xu and W. W. Webb, J. Opt. Soc. Am. B 13, 481 (1996).
  50. M. Balu, T. Baldacchini, J. Carter, T. B. Krasieva, R. Zadoyan, and B. J. Tromberg, J. Biomed. Opt. 14, 010508 (2009).
  51. H. Guo, K.-I. Sato, K. Takashima, and H. Yokoyama, in Proceedings of International Conference on Ultrafast Phenomena 2006 TuE8 (2006).
  52. H. Yokoyama, H. Guo, T. Yoda, K. Takashima, K. Sato, H. Taniguchi, and H. Ito, Opt. Express 14, 3467 (2006).
  53. M. Kuramoto, N. Kitajima, H. Guo, Y. Furushima, M. Ikeda, and H. Yokoyama, Opt. Lett. 32, 2726 (2007).
  54. S. W. Chu, I. H. Chen, T. M. Liu, P. C. Chen, C. K. Sun, and B. L. Lin, Opt. Lett. 26, 1909 (2001).
  55. C. Wang, L. Qiao, F. He, Y. Cheng, and Z. Xu, J. Microsc-Oxford 243, 179 (2011).
  56. C. Wang, L. L. Qiao, Z. L. Mao, Y. Cheng, and Z. Z. Xu, J. Opt. Soc. Am. B 25, 976 (2008).
  57. K. Murari, Y. Y. Zhang, S. P. Li, Y. P. Chen, M. J. Li, and X. D. Li, Opt. Lett. 36, 1299 (2011).
  58. S. Yazdanfar, C. Joo, C. Zhan, M. Y. Berezin, W. J. Akers, and S. Achilefu, J. Biomed. Opt. 15, 030505 (2010).
  59. H. Yokoyama, H. Tsubokawa, H. Guo, J. Shikata, K. Sato, K. Takashima, K. Kashiwagi, N. Saito, H. Taniguchi, and H. Ito, J. Biomed. Opt. 12, 054019 (2007).
  60. K. Taira, T. Hashimoto, and H. Yokoyama, Opt. Express 15, 2454 (2007).
  61. J. R. Unruh, E. S. Price, R. G. Molla, L. Stehno-Bittel, C. K. Johnson, and R. Hui, Opt. Express 14, 9825 (2006).
  62. R. Aviles-Espinosa, Y. Ding, M. A. Cataluna, D. Nikitichev, P. Loza-Alvarez, and E. Rafailov, in Proceedings of CLEO: Applications and Technology ATh1M.2 (2012).
  63. G. D. Reddy and P. Saggau, J. Biomed. Opt. 10, 064038 (2005).
  64. R. H. Jiang, X. H. Lu, D. R. Li, T.W. Quan, X. L. Liu, Q. M. Luo, and S. Q. Zeng, Prog. Biochem. Biophys. 39, 505 (2012).
  65. K. H. Kim, C. Buehler, K. Bahlmann, T. Ragan, W. C. Lee, E. Nedivi, E. L. Heffer, S. Fantini, and P. T. So, Opt. Express 15, 11658 (2007).
  66. S. S. Howard, A. A. Straub, and C. Xu, in Proceedings of CLEO CWD6 (2010).
  67. A. A. Straub, S. S. Howard, and C. Xu, in Proceedings of CLEO: Applications and Technology ATh4C.3 (2012).
  68. D. Oron, E. Tal, and Y. Silberberg, Opt. Express 13, 1468 (2005).
  69. J. Y. Yu, C. H. Kuo, D. B. Holland, Y.Chen,M. Ouyang, G. A. Blake, R. Zadoyan, and C. L. Guo, J. Biomed. Opt. 16, 116009 (2011).
  70. T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, Nature Methods 8, 417 (2011).
  71. P. Kim, M. Puoris'haag, D. Cote, C. P. Lin, and S. H. Yun, J. Biomed. Opt. 13, 010501 (2008).
  72. J. C. Jung and M. J. Schnitzer, Opt. Lett. 28, 902 (2003).
  73. G. J. Liu, T. Q. Xie, I. V. Tomov, J. P. Su, L. F. Yu, J. Zhang, B. J. Tromberg, and Z. P. Chen, Opt. Lett. 34, 2249 (2009).
  74. W. Gobel, J. N. Kerr, A. Nimmerjahn, and F. Helmchen, Opt. Lett. 29, 2521 (2004).
  75. M. J. Levene, D. A. Dombeck, K. A. Kasischke, R. P. Molloy, andW.W.Webb, J. Neurophys. 91, 1908 (2004).
  76. W. Piyawattanametha, R. P. Barretto, T. H. Ko, B. A. Flusberg, E. D. Cocker, H. Ra, D. Lee, O. Solgaard, and M. J. Schnitzer, Opt. Lett. 31, 2018 (2006).
  77. D. R. Rivera, C. M. Brown, D. G. Ouzounov, I. Pavlova, D. Kobat, W. W. Webb, and C. Xu, Proc. Natl. Acad. Sci. USA 108, 17598 (2011).
  78. C. L. Hoy, N. J. Durr, P. Y. Chen, W. Piyawattanametha, H. Ra, O. Solgaard, and A. Ben-Yakar, Opt. Express 16, 9996 (2008).
  79. Y. Wu, Y. Leng, J. Xi, and X. Li, Opt. Express 17, 7907 (2009).
  80. Y. Y. Zhang, M. L. Akins, K. Murari, J. F. Xi, M. J. Li, K. Luby-Phelps, M. Mahendroo, and X. D. Li, Proc. Natl. Acad. Sci. USA 109, 12878 (2012).
  81. W. Denk, Proc. Natl. Acad. Sci. USA 91, 6629 (1994).
  82. K. A. Kasischke, H. D. Vishwasrao, P. J. Fisher, W. R. Zipfel, and W. W. Webb, Science 305, 99 (2004).
  83. H. D. Vishwasrao, A. A. Heikal, K. A. Kasischke, and W. W. Webb, J. Biol. Chem. 280, 25119 (2005).
  84. C. Stosiek, O. Garaschuk, K. Holthoff, and A. Konnerth, Proc. Natl. Acad. Sci. USA 100, 7319 (2003).
  85. A. Sobczyk, V. Scheuss, and K. Svoboda, J. Neurosci. 25, 6037 (2005).
  86. A. Cheng, J. T. Goncalves, P. Golshani, K. Arisaka, and C. Portera-Cailliau, Nat. Methods 8, 139 (2011).
  87. W. Zheng, D. Li, Y. Zeng, Y. Luo, and J. Y. Qu, Biomed. Opt. Express 2, 71 (2010).
  88. D. Li, W. Zheng, Y. Zeng, Y. Luo, and J. Y. Qu, Opt. Lett. 36, 834 (2011).
  89. M. A. DeBernardi and G. Brooker, Method Enzymol. 414, 317 (2006).
  90. B. C. Dickinson, J. Peltier, D. Stone, D. V. Schaffer, and C. J. Chang, Nat. Chem. Biol. 7, 106 (2011).
  91. J. R. Stone and S. Yang, Antioxid Redox Signal 8, 243 (2006).
  92. M. A. Go, C. Stricker, S. Redman, H.-A. Bachor, and V. R. Daria, J. Biophoton. 5, 745 (2012).
  93. N. Nishimura, C. B. Schaffer, B. Friedman, P. S. Tsai, P. D. Lyden, and D. Kleinfeld, Nat. Methods 3, 99 (2006).
  94. J. Bosch, S. Yusuf, J. Pogue, P. Sleight, E. Lonn, B. Rangoonwala, R. Davies, J. Ostergren, J. Probstfield, and H. Investigators, Brit. Med. J. 324, 699 (2002).
  95. S. Cho, E. M. Park, M. Febbraio, J. Anrather, L. Park, G. Racchumi, R. L. Silverstein, and C. Iadecola, J. Neurosci. 25, 2504 (2005).
  96. Y. Bao, L. Y. Qin, E. Kim, S. Bhosle, H. C. Guo, M. Febbraio, R. E. Haskew-Layton, R. Ratan, and S. Cho, J. Cerebr. Blood Flow Met. 32, 1567 (2012).
  97. H. Ujike, M. Takaki, M. Kodama, and S. Kuroda, Ann. Ny. Acad. Sci. 965, 55 (2002).
  98. T. H. Chia and M. J. Levene, J. Neurophys. 102, 1310 (2009).
  99. S. Zhuo, X. Zhu, G. Wu, J. Chen, and S. Xie, J. Biomed. Opt. 16, 120501 (2011).
  100. S. Zhuo, J. Yan, G. Chen, H. Shi, X. Zhu, J. Lu, J. Chen, and S. Xie, Plos One 7, e38655 (2012).
  101. M. Elangovan, H. Wallrabe, Y. Chen, R. N. Day, M. Barroso, and A. Periasamy, Methods 29, 58 (2003).
  102. K. M. Berland, P. T. C. So, and E. Gratton, Biophys. J. 68, 694 (1995).
  103. S. A. Kim, K. G. Heinze, and P. Schwille, Nat Methods 4, 963 (2007).
  104. R. Yasuda, C. D. Harvey, H. Zhong, A. Sobczyk, L. van Aelst, and K. Svoboda, Nat. Neurosci. 9, 283 (2006).
  105. Y. E. Chen and A. Periasamy, Microsc. Res. Techniq. 63, 72 (2004).
  106. A. Jeshtadi, P. Burgos, C. D. Stubbs, A. W. Parker, L. A. King, M. A. Skinner, and S. W. Botchway, J. Virol 84, 12886 (2010).
  107. K. Kucharz, T. Wieloch, and H. Toresson, J. Cereb Blood Flow Met. 31, 1663 (2011).
  108. G. Moneron and S. W. Hell, Opt. Express 17, 14567 (2009).
  109. P. Bianchini and A. Diaspro, J. Microsc-Oxford 245, 225 (2012).
  110. N. Jia, T. R. Sato, and E. Betzig, Proc. Natl. Acad. Sci. USA 109, 22 (2012).
  111. N. Ji, D. E. Milkie, and E. Betzig, Nat. Methods 7, 141 (2010).
  112. E. Papagiakoumou, F. Anselmi, A. Begue, V. de Sars, J. Gluckstad, E. Y. Isacoff, and V. Emiliani, Nat. Methods 7, 848 (2010).
  113. B. K. Andrasfalvy, B. V. Zemelman, J. Tang, and A. Vaziri, Proc. Natl. Acad. Sci. USA 107, 11981 (2010).
  114. J. P. Rickgauer and D. W. Tank, Proc. Natl. Acad. Sci. USA 106, 15025 (2009).
  115. S. A. Yuan, C. A. Roney, J. Wierwille, C. W. Chen, B. Y. Xu, G. Griffiths, J. Jiang, H. Z. Ma, A. Cable, R. M. Summers, and Y. Chen, Phys. Med. Biol. 55, 191 (2010).
  116. J. Xi, K. Murari, Y. Zhang, Y. Chen, J. Li, and X. Li, in Proceedings of Bio-Optics: Design and Application BWA5 (2011).
  117. B. Jeong, B. Lee, M. S. Jang, H. Nam, S. J. Yoon, T. Wang, J. Doh, B. G. Yang, M. H. Jang, and K. H. Kim, Opt. Express 19, 13089 (2011).
  118. C. Vinegoni, T. Ralston, W. Tan, W. Luo, D. L. Marks, and S. A. Boppart, Appl. Phys. Lett. 88, 053901 (2006).
  119. S. Tang, C. H. Sun, T. B. Krasieva, Z. P. Chen, and B. J. Tromberg, Opt. Lett. 32, 503 (2007).
  120. J. Kneipp, H. Kneipp, and K. Kneipp, Proc. Natl. Acad. Sci. USA 103, 17149 (2006).
  121. C. H. Chien, W. W. Chen, J. T. Wu, and T. C. Chang, J. Biomed. Opt. 16, 016012 (2011).

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