Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo
Noninvasive harmonics optical microscopy for long-term observation of embryonic nervous system development in vivo
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DOI：
10.1117/1.2363369
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发表时间：
2006-09-01
影响因子：
3.5
通讯作者：
Sun, Chi-Kuang
中科院分区：
文献类型：
--
作者：
Chen, Szu-Yu;Hsieh, Cho-Shuen;Sun, Chi-Kuang
神经系统发育是一个复杂的动态过程，许多机制尚不清楚。利用内源性二次谐波产生作为极化神经纤维的对比，利用三次谐波产生（THG）揭示形态学变化，我们成功地在1230 nm光源下观察了脊椎动物胚胎神经发育的全过程。在没有荧光标记的情况下，可以连续记录活斑马鱼胚胎内神经系统的动态发育超过20小时。由于THG过程不像基于荧光的技术那样受到基因表达和分化时间的限制，因此可观察到的阶段可以提前到发育过程的最开始。从神经板到神经管的完整三维大脑发育可以用亚微米的横向分辨率来揭示。我们还首次报道了从有髓神经纤维和具有堆叠膜结构的光感受器的外段产生SHG。我们的研究清楚地表明，基于高次谐波的光学显微镜具有很强的潜力，长期在体内研究神经系统，包括神经系统的遗传疾病，轴突寻路，神经再生，神经修复，神经修复和神经干细胞发育。(c)2006年，由光学仪器工程师协会（Society of Photo-Optical Instrumentation Engineers）主办。
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 repair, and neural stem cell development. (c) 2006 Society of Photo-Optical Instrumentation Engineers.