期刊
JOURNAL OF BIOMEDICAL OPTICS
卷 13, 期 6, 页码 -出版社
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.3050423
关键词
in vivo imaging; harmonic generation microscopy; third harmonic generation; second harmonic generation; three-photon excited fluorescence; embryology; gene expression; morphant technology
资金
- National Health Research Institute [NHRI-EX97-9201EI]
- National Science Council [NSC-96-2313-B-019-004, NSC-95-2313-B019-010]
- National Taiwan University
- National Taiwan University Center for Medical Excellence
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-1 alpha, 2 alpha, and 3 alpha 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. (c) 2008 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3050423]
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