Long-term in vivo study of vertebrate embryonic development using noninvasive harmonics optical microscopy

Szu-Yu Chen; C.-S. Hsieh; S.-W. Chu; Cheng-Yung Lin; C.-Y. Ko; Y.-C. Chen; Huai-Jen Tsai; C.-H. Hu; Chi-Kuang Sun

doi:10.1117/12.590169

30 March 2005 Long-term in vivo study of vertebrate embryonic development using noninvasive harmonics optical microscopy

Szu-Yu Chen, C.-S. Hsieh, S.-W. Chu, Cheng-Yung Lin, C.-Y. Ko, Y.-C. Chen, Huai-Jen Tsai, C.-H. Hu, Chi-Kuang Sun

Author Affiliations +

Proceedings Volume 5700, Multiphoton Microscopy in the Biomedical Sciences V; (2005) https://doi.org/10.1117/12.590169
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

Harmonics optical microscopy (HOM) provides a truly “noninvasive” tool for in vivo and long-term study of vertebrate embryonic development. Based on the nonlinear natures, it provides sub-micrometer 3D spatial resolution and high 3D optical-sectioning power (~1μm axial resolution) without using invasive and toxic fluorophores. Since only virtual-level-transition is involved, HOM is known to leave no energy deposition and no photodamages. Combined with second harmonic generation, which is sensitive to specific structure such as nerve and muscle fibers, HOM can be used to do 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. Zebrafish embryos now have been used to study many vertebrate physiological systems. We have demonstrated an in vivo HOM study of developmental dynamics of several 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.

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Szu-Yu Chen, C.-S. Hsieh, S.-W. Chu, Cheng-Yung Lin, C.-Y. Ko, Y.-C. Chen, Huai-Jen Tsai, C.-H. Hu, and Chi-Kuang Sun "Long-term in vivo study of vertebrate embryonic development using noninvasive harmonics optical microscopy", Proc. SPIE 5700, Multiphoton Microscopy in the Biomedical Sciences V, (30 March 2005); https://doi.org/10.1117/12.590169

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KEYWORDS

Second-harmonic generation

In vivo imaging

Nerve

Optical fibers

Brain

Tissues

Standards development

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