Research Papers

Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo

[+] Author Affiliations
Szu-Yu Chen, Cho-Shuen Hsieh, Shi-Wei Chu

National Taiwan University, Department of Electrical Engineering and Graduate Institute of Electro-Optical Engineering, Taipei 10617, Taiwan

Cheng-Yung Lin

National Taiwan University, Institute of Molecular and Cellular Biology, Taipei 10617, Taiwan

Ching-Yi Ko, Yi-Chung Chen

National Taiwan Ocean University, Institute of Bioscience and Biotechnology, Keelung 20224, Taiwan

Huai-Jen Tsai

National Taiwan University, Institute of Molecular and Cellular Biology, Taipei 10617, Taiwan

Chin-Hwa Hu

National Taiwan Ocean University, Institute of Bioscience and Biotechnology, Keelung 20224, Taiwan

Chi-Kuang Sun

National Taiwan University, Department of Electrical Engineering and Graduate Institute of Electro-Optical Engineering, Taipei 10617, Taiwan

J. Biomed. Opt. 11(5), 054022 (October 30, 2006). doi:10.1117/1.2363369
History: Received October 18, 2005; Revised March 20, 2006; Accepted May 05, 2006; Published October 30, 2006; Online October 30, 2006
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Nervous system development is a complicated dynamic process, and many mechanisms remain unknown. By utilizing endogenous second-harmonic-generation as the contrast of polarized nerve fibers and third-harmonic-generation (THG) to reveal morphological changes, we have successfully observed the vertebrate embryonic nervous development from the very beginning based on a 1230-nm light source. The dynamic development of the nerve system within a live zebrafish embryo can be recorded continuously more than 20hr without fluorescence markers. Since the THG process is not limited by the time of gene expression and differentiation as fluorescence-based techniques are, the observable stages can be advanced to the very beginning of the development process. The complete three-dimensional brain development from a neural plate to a neural tube can be uncovered with a submicron lateral resolution. We have, for the first time, also reported the generation of SHG from myelinated nerve fibers and the outer segment of the photoreceptors with a stacked membrane structure. Our study clearly indicates the fact that higher-harmonics-based optical microscopy has the strong potential to long-term in vivo study of the nervous system, including genetic disorders of the nervous system, axon pathfinding, neural regeneration, neural repair, and neural stem cell development.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

Szu-Yu Chen ; Cho-Shuen Hsieh ; Shi-Wei Chu ; Cheng-Yung Lin ; Ching-Yi Ko, et al.
"Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo", J. Biomed. Opt. 11(5), 054022 (October 30, 2006). ; http://dx.doi.org/10.1117/1.2363369


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