Higher harmonic generation microscopy for developmental biology
Resource
Journal of Structural Biology 147 (1): 19-30
Journal
Journal of Structural Biology
Journal Volume
147
Journal Issue
1
Pages
19-30
Date Issued
2004
Author(s)
Sun, CK
Chu, SW
Chen, Szu-Yu
Chen, SY
Tsai, Tsung-Han
Tsai, TH
Liu, TM
Liu, Tzu-Ming
Lin, CY
Lin, Chung-Yung
Tsai, Huai-Jen
Tsai, HJ
Abstract
Optical higher harmonic generation, including second harmonic generation and third harmonic generation, 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 (3D) 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 sub-cellular 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. © 2003 Elsevier Inc. All rights reserved.
Subjects
Developmental biology; Embryo; Higher harmonic generation; Multi-dimensional microscopy; Scanning microscopy; Second harmonic generation; Third harmonic generation
Other Subjects
animal tissue; article; controlled study; cytoarchitecture; ectogenesis; embryo; embryology; energy conservation; illumination; image analysis; larval stage; multiphoton microscopy; nonhuman; optical density; priority journal; scanning electrochemical microscopy; zebra fish; Animals; Developmental Biology; Embryo, Nonmammalian; Imaging, Three-Dimensional; Lasers; Mathematics; Microscopy; Zebrafish; Danio; Danio rerio
Type
journal article