Second Harmonic Generation Microscopy Versus Third Harmonic Generation Microscopy in Biological Tissues

  • Chi-Kuang Sun
Part of the Springer Series in Optical Sciences book series (SSOS, volume 87)


Second-harmonic generation (SHG) and third-harmonic generation (THG) processes are both nonlinear processes, related to the interaction of intense light with matters. SHG process describes the generation of light wave that is twice the frequency (with half of the original wavelength) of the original one while THG process describes the generation of light wave that triples the frequency (with one third of the original wavelength) of the original one. The harmonic light wave generation is coupled from the excited nonlinear polarization P NL under intense laser excitation. The interaction of nonlinear polarization P NL and the excitation light is usually related through a nonlinear susceptibility χ, as previously described in Chaps. 7 and 8. SHG and THG can be visualized by considering the interaction in terms of the exchange of photons between various frequencies of the fields. According to this picture, which is previously illustrated in Figs. 8.1 (a), (b), two or three photons of angular frequency ω are destroyed and a photon of angular frequency 2ω (for SHG) or 3ω (for THG) is created in a single quantum-mechanical process. The solid lines in the figure represent the atomic ground states, and the dashed lines represent what are known as virtual levels. These virtual levels are not energy eigenlevels of the atoms, but rather represent the combined energy of one of the energy eigenstates of the atom and one or more photons of the radiation field. Due to its virtual level transition characteristics, harmonic generations are known to leave no energy deposition to the interacted matters, since no real transition involved and the emitted photon energy will be exactly the same as the total absorbed photon energy. This virtual transition characteristic provides the optical “noninvasive” nature desirable for microscopy applications, especially for live biological imaging.


Starch Granule Purple Membrane Nonlinear Photonic Crystal Forsterite Laser Maize Stem 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    R. Hellwarth and P. Christensen: Opt. Commun. 12, 318 (1974)ADSCrossRefGoogle Scholar
  2. 2.
    C. J. R. Sheppard, R. Kompfner, J. Gannaway, and D. Walsh: IEEE J. Quantum Electron. 13, 100 (1977)Google Scholar
  3. 3.
    G. T. Boyd, Y. R. Shen, and T. W. H¨ansch: Opt. Lett. 11, 97 (1986)ADSCrossRefGoogle Scholar
  4. 4.
    Y. Uesu, S. Kurimura, and Y. Yamamoto: Appl. Phys. Lett. 66, 2165 (1995)ADSCrossRefGoogle Scholar
  5. 5.
    R. Gauderon, P. B. Lukins, and C. J. R. Sheppard: Opt. Lett. 23, 1209 (1998)ADSCrossRefGoogle Scholar
  6. 6.
    We follow the notation of H. A. Haus: Waves and Fields in Optoelectronics (Prentice-Hall, Inc., New Jersey 1984 )Google Scholar
  7. 7.
    C.-K. Sun, S.-W. Chu, S.-P. Tai, S. Keller, U. K. Mishra, and S. P. DenBaars: Appl. Phys. Lett. 77, 2331 (2000)ADSCrossRefGoogle Scholar
  8. 8.
    C.-K. Sun, S.-W. Chu, S. P. Tai, S. Keller, A. Abare, U. K. Mishra, and S. P. DenBaars: J. Scanning Microscopies 23, 182 (2001)CrossRefGoogle Scholar
  9. 9.
    F. Bresson, F. E. Hernandez, J.-W. Shi, and C.-K. Sun: ‘Electric-field induced second harmonic generation in nematic liquid crystals as a probe for electric field’, In: Proceeding of Optics and Photonics Taiwan ‘01, ed. by (Kaohsiung, Taiwan ROC 2001 ) pp. 713–715Google Scholar
  10. 10.
    F. Bresson, C.C. Chen, F.E. Hernandez, J.-W. Shi, and C.-K. Sun: submitted to Rev. Sci. Instrum.Google Scholar
  11. 11.
    S. Fine and W. P. Hansen: Appl. Optics 10, 2350 (197 1)Google Scholar
  12. 12.
    I. Freund, M. Deutsch, and A. Sprecher: Biophys. J. 50, 693 (1986)CrossRefGoogle Scholar
  13. 13.
    J. Y. Huang, Z. Chen, and A. Lewis: J. Phys. Chem. 93, 3314 (1989)CrossRefGoogle Scholar
  14. 14.
    Y. Guo, P. P. Ho, A. Tirksliunas, F. Liu, and R. R. Alfano: Appl. Optics 35, 6810 (1996)ADSCrossRefGoogle Scholar
  15. 15.
    Y. Guo, P. P. Ho, H. Savage, D. Harris, P. Sacks, S. Schantz, F. Liu, N. Zhadin, and R. R. Alfano: Opt. Lett. 22, 1323 (1997)ADSCrossRefGoogle Scholar
  16. 16.
    O. Bouevitch, A. Lewis, I. Pinevsky, J. P. Wuskell, and L. M. Loew: Biophys. J. 65, 672 (1993)CrossRefGoogle Scholar
  17. 17.
    P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew: Biophys. J. 77, 3341 (1999)CrossRefGoogle Scholar
  18. 18.
    G. Peleg, A. Lewis, M. Linial, and L. M. Loew: P. Natl. Acad. Sci. USA 96, 6700 (1999)ADSCrossRefGoogle Scholar
  19. 19.
    L. Moreax, O. Sandre, and J. Mertz: J. Opt. Soc. Am. B 17, 1685 (2000)ADSCrossRefGoogle Scholar
  20. 20.
    F.-J. Kao, Y.-S. Wang, M.-K. Huang, S. L. Huang, P.-C. Cheng: P. Soc. Photo–Opt. Inst. 4082, 119 (2000)Google Scholar
  21. 21.
    T. Zhao, Z.-H. Chen, F. Chen, W.-S. Shi, H.-B. Lu, and G.-Z. Yang: Phys. Rev. B 60, 1697 (1999)ADSCrossRefGoogle Scholar
  22. 22.
    Y. Dumeige, P. Vidakovic, S. Sauvage, I. Sagnes, J. A. Levenson, C. Sibilia, M. Centini, G. D’Aguanno, and M. Scalora: Appl. Phys. Lett. 78, 3021 (2001)ADSCrossRefGoogle Scholar
  23. 23.
    V. Berger: Phys. Rev. Lett. 81, 4136 (1998)ADSCrossRefGoogle Scholar
  24. 24.
    N. G. R. Broderick, G. W. Ross, H. L. Offerhaus, D. J. Richardson, and D. C. Hanna: Phys. Rev. Lett. 84, 4345 (2000)ADSCrossRefGoogle Scholar
  25. 25.
    K. Clays. S. Van Elshocht, M. Chi, E. Lepoudre, and A. Persoons: J. Opt. Soc. Am. B 18, 1474 (2001)ADSCrossRefGoogle Scholar
  26. 26.
    S.-W. Chu, I-H. Chen, T.-M. Liu, B.-L. Lin, P. C. Cheng, and C.-K. Sun: Opt. Lett. 26, 1909 (2001)ADSCrossRefGoogle Scholar
  27. 27.
    T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson: J. Scanning Microscopies 23, 249 (2001)CrossRefGoogle Scholar
  28. 28.
    R. R. Anderson and J. A. Parish: J. Invest. Dermat. 77, 13 (198 1)Google Scholar
  29. 29.
    P. C. Cheng, S. J. Pan, A. Shih, K.-S. Kim, W. S. Liou, and M. S. Park: J. Microscopy 189, 199 (1998)CrossRefGoogle Scholar
  30. 30.
    B. E. Bouma, G. J. Tearney, I. P. Bilinsky, B. Golubovic, and J. G. Fujimoto: Opt. Lett. 21, 1839 (1996)ADSCrossRefGoogle Scholar
  31. 31.
    D. J. Gallant, B. Bouchet, and P. M. Baldwin: Carbohydrate Polymers 32, 177 (1997)CrossRefGoogle Scholar
  32. 32.
    T. Y. F. Tsang: Phys. Rev. A 52, 4116 (1995)ADSCrossRefGoogle Scholar
  33. 33.
    R. W. Boyd: Nonlinear Optics ( Academic Press, San Diego, CA 1992 )Google Scholar
  34. 34.
    Y. Barad, H. Eisenberg, M. Horowitz, and Y. Silberberg: Appl. Phys. Lett. 70, 922 (1997)ADSCrossRefGoogle Scholar
  35. 35.
    M. Muller, J. Squier, K. R. Wilson, and G. J. Brakenhoff: J. Microscopy 191, 266 (1998)CrossRefGoogle Scholar
  36. 36.
    J. A. Squier, M. Muller, G. J. Brakenhoff, and K. R. Wilson: Opt. Express 3, 315 (1998)ADSCrossRefGoogle Scholar
  37. 37.
    D. Yelin and Y. Silberberg: Opt. Express 5, 169 (1999)ADSCrossRefGoogle Scholar
  38. 38.
    J. A. Squier and M. Muller: Appl. Optics 38, 5789 (1999)ADSCrossRefGoogle Scholar
  39. 39.
    D. Yelin, Y. Silberberg, Y. Barad, and J. S. Patel: Appl. Phys. Lett. 74, 3107 (1999)ADSCrossRefGoogle Scholar
  40. 40.
    L. Canioni, S. Rivet, L. Sarger, R. Barille, P. Vacher, and P. Voisin: Opt. Lett. 26, 515 (2001)ADSCrossRefGoogle Scholar
  41. 41.
    I-H. Chen, S.-W. Chu, C.-K. Sun, P. C. Cheng, and B.-L. Lin: ‘Wavelength dependent cell damages in multi-photon confocal microscopy’. In: Optical and Quantum Electronics (in press 2002)Google Scholar
  42. 42.
    T.-M. Liu, S.-W. Chiu, I-H. Chen, C.-K. Sun, B.-L. Lin, W.-J. Yang, P.-C. Cheng, and I. Johnson: ‘Multi-photon fluorescence of green fluorescence protein (GFP) and commonly used bio-probes excited by femtosecond Cr:forsterite lasers’, In: Proceedings of 2nd International Photonics Conference, ed. by Hsinchu (TAIWAN, 2000 119) pp. 121–Google Scholar
  43. 43.
    T.-M. Liu, S.-W. Chu, C.-K. Sun, B.-L. Lin, P. C. Cheng, and I. Johnson: J. Scanning Microscopies 23, 249 (2001)CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2003

Authors and Affiliations

  • Chi-Kuang Sun

There are no affiliations available

Personalised recommendations