Long-term in vivo harmonics imaging of zebrafish embryonic development based on a femtosecond Cr:forsterite laser

S. Y. Chen, T. H. Tsai, C. S. Hsieh, S. P. Tai, C. Y. Lin, C. Y. Ko, Y. C. Chen, H. J. Tsai, C. H. Hu, C. K. Sun

Research output: Contribution to journalConference articlepeer-review


Based on a femtosecond Cr:forsterite laser, harmonics optical microscopy (HOM) provides a truly "noninvasive" tool for in vivo and long-term study of vertebrate embryonic development. Based on optial nonlinearity, HOM provides submicrometer 3D spatial resolution and high 3D optical-sectioning power without using invasive and toxic fluorophores. Since only virtual-level-transition is involved, HOM is known to leave no energy deposition and no photodamage. Combined with second harmonic generation, which is sensitive to specific structure such as nerve and muscle fibers, HOM can perform functional studies of early developmental dynamics of many vertebrate physiological systems. Recently, zebrafish has become a standard model for many biological and medical studies of vertebrates, due to the similarity between embryonic development of zebrafish and human being. Here we demonstrate hi vivo HOM studies of developmental dynamics of several important embryonic physiological systems in live zebrafish embryos, with focuses on the developments of brains, eyes, ears, and hearts. Based on a femtosecond Cr:forsterite laser, which provides the deepest penetration (∼1.5mm) and least photodamage in the zebrafish embryo, complete developing processes of different physiological systems within a period of time longer than 20 hours can be non-invasively observed inside the same embryo.

Original languageEnglish
Article number14
Pages (from-to)73-80
Number of pages8
JournalProceedings of SPIE - The International Society for Optical Engineering
StatePublished - 2005
EventCommercial and Biomedical Applications of Ultrafast Lasers V - San Jose, CA, United States
Duration: 24 Jan 200527 Jan 2005


  • Development
  • Femtosecond laser
  • Harmonic generation
  • In vivo
  • Noninvasive
  • Nonlinear optics
  • Zebrafish


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