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Higher harmonic generation microscopy for developmental biology
Chi-Kuang Sun1; Shi-Wei Chu1; Szu-Yu Chen1; Tsung-Han Tsai1; Tzu-Ming Liu1; Chung-Yung Lin2; Huai-Jen Tsai2
Source PublicationJournal of Structural Biology

Optical higher harmonic generation, including second harmonic generation and third harmonicgeneration, leaves no energy deposition to its interacted matters due to an energy-conservation characteristic, providing the "noninvasiveness" nature desirable for biological studies. Combined with its nonlinearity, higher harmonic generation microscopy provides excellent three-dimensional (313) sectioning capability, offering new insights into the studies of embryonic morphological changes and complex developmental processes. By choosing a laser working in the biological penetration window, here we present a noninvasive in vivo light microscopy with sub-micron 3D resolution and millimeter penetration, utilizing endogenous higher harmonic generation signals in live specimens. Noninvasive imaging was performed in live zebrafish (Danio rerio) embryos. The complex developmental processes within > 1 mm-thick zebrafish embryos can be observed in vivo without any treatment. No optical damage was found even with high illumination after long-term observations and the examined embryos all developed normally at least to the larval stage. The excellent 3D resolution of the demonstrated technology allows us to capture the subtle developmental information on the cellular or subcellular levels occurring deep inside the live embryos and larvae. This technique can not only provide in vivo observation of the cytoarchitecture dynamics during embryogenesis with submicron resolution and millimeter penetration depth, but would also make strong impact in developmental and structural biology studies. (C) 2003 Elsevier Inc. All rights reserved.

KeywordHigher Harmonic Generation Third Harmonic Generation Second Harmonic Generation Embryo Scanning Microscopy Developmental Biology Multi-dimensional Microscopy
Indexed BySCIE
WOS Research AreaBiochemistry & Molecular Biology ; Biophysics ; Cell Biology
WOS SubjectBiochemistry & Molecular Biology ; Biophysics ; Cell Biology
WOS IDWOS:000221246000004
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Cited Times [WOS]:152   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.Graduate Institute of Electro-Optical Engineering and Department of Electrical Engineering, National Taiwan University, Taipei 10617, Taiwan, ROC
2.Institute of Molecular and Cell Biology, National Taiwan University, Taipei 10617, Taiwan, ROC
Recommended Citation
GB/T 7714
Chi-Kuang Sun,Shi-Wei Chu,Szu-Yu Chen,et al. Higher harmonic generation microscopy for developmental biology[J]. Journal of Structural Biology,2004,147(1):19-30.
APA Chi-Kuang Sun,Shi-Wei Chu,Szu-Yu Chen,Tsung-Han Tsai,Tzu-Ming Liu,Chung-Yung Lin,&Huai-Jen Tsai.(2004).Higher harmonic generation microscopy for developmental biology.Journal of Structural Biology,147(1),19-30.
MLA Chi-Kuang Sun,et al."Higher harmonic generation microscopy for developmental biology".Journal of Structural Biology 147.1(2004):19-30.
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