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Journal of Biomedical Optics

In vivo long-term continuous observation of gene expression in zebrafish embryo nerve systems by using harmonic generation microscopy and morphant technology
Author(s): Cho-Shuen Hsieh; Ching-Yi Ko; Szu-Yu Chen; Tzu-Ming Liu; Jian-Shiung Wu; Chin-Hwa Hu; Chi-Kuang Sun
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Paper Abstract

Gene expression plays an important role in embryo development and organ function. Previous studies have shown that harmonic generation microscopy (HGM) can be used as a fluorescence signal-independent, minimally invasive method with a subcellular 3-D resolution and a penetration depth in the order of millimeters for long-term continuous imaging of vertebrate embryos. We show that it is ideal to combine in vivo HGM with the morphant technology for minimally invasive, long-term continuous observation of gene expression in the nervous system of vertebrate embryos. Since second- and third-harmonic generations (SHG, THG) are virtual-state-transition-based systems that depend only on the structure of the organisms, they are not temporally limited by the expression of the fluorescence proteins. We successfully identified the expression of the zarnt2a and the hif-1α, 2α, and 3α genes in the nervous system of zebrafish embryos with specific knockdown genes by microscopically observing the embryos from the early stages of embryogenesis. The results from a combination of the two different modalities, i.e., SHG microscopy and THG microscopy, successfully revealed the weak cell adhesion, cell apoptosis, nerve formation reduction, and neural tube distortion in the morphant zebrafish embryos.

Paper Details

Date Published: 1 November 2008
PDF: 7 pages
J. Biomed. Opt. 13(6) 064041 doi: 10.1117/1.3050423
Published in: Journal of Biomedical Optics Volume 13, Issue 6
Show Author Affiliations
Cho-Shuen Hsieh, National Taiwan Univ. (Taiwan)
Ching-Yi Ko, National Taiwan Ocean Univ. (Taiwan)
Szu-Yu Chen, National Taiwan Univ. (Taiwan)
Tzu-Ming Liu, National Taiwan Univ. (Taiwan)
Jian-Shiung Wu, National Taiwan Ocean Univ. (Taiwan)
Chin-Hwa Hu, National Taiwan Ocean Univ. (Taiwan)
Chi-Kuang Sun, National Taiwan Univ. (Taiwan)

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