PUBLICATION

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

Authors
Chen, S.Y., Hsieh, C.S., Chu, S.W., Lin, C.Y., Ko, C.Y., Chen, Y.C., Tsai, H.J.,Hu, C.H., and Sun, C.K.
ID
ZDB-PUB-061116-4
Date
2006
Source
Journal of Biomedical Optics   11(5): 54022 (Journal)
Registered Authors
Hu, Chin-Hwa, Tsai, Huai-Jen
Keywords
none
MeSH Terms
  • Animals
  • Brain/cytology*
  • Brain/embryology*
  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement/instrumentation*
  • Image Enhancement/methods
  • Microscopy, Confocal/instrumentation*
  • Microscopy, Confocal/methods
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Zebrafish/anatomy & histology*
  • Zebrafish/embryology*
PubMed
17092171 Full text @ J. Biomed. Opt.
Abstract
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 20 hr 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.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes