Filippidis George, Kouloumentas Christos, Voglis Giannis, Zacharopoulou Fotini, Papazoglou Theodore G, Tavernarakis Nektarios
Foundation of Research and Technology-Hellas, Institute of Electronic Structure and Laser, PO Box 1527, Heraklion, Greece 71110.
J Biomed Opt. 2005 Mar-Apr;10(2):024015. doi: 10.1117/1.1886729.
Second-harmonic generation (SHG) and two-photon excitation fluorescence (TPEF) are relatively new and promising tools for the detailed imaging of biological samples and processes at the microscopic level. By exploiting these nonlinear phenomena phototoxicity and photobleaching effects on the specimens are reduced dramatically. The main target of this work was the development of a compact inexpensive and reliable experimental apparatus for nonlinear microscopy measurements. Femtosecond laser pulses were utilized for excitation. We achieved high-resolution imaging and mapping of Caenorhabditis elegans (C. elegans) neurons and muscular structures of the pharynx, at the microscopic level by performing SHG and TPEF measurements. By detecting nonlinear phenomena such as SHG and TPEF it is feasible to extract valuable information concerning the structure and the function of nematode neurons.
二次谐波产生（SHG）和双光子激发荧光（TPEF）是相对较新且很有前景的工具，可用于在微观层面详细成像生物样本和过程。通过利用这些非线性现象，对样本的光毒性和光漂白效应会大幅降低。这项工作的主要目标是开发一种用于非线性显微镜测量的紧凑、廉价且可靠的实验装置。利用飞秒激光脉冲进行激发。通过执行SHG和TPEF测量，我们在微观层面实现了秀丽隐杆线虫（C. elegans）神经元以及咽部肌肉结构的高分辨率成像和映射。通过检测诸如SHG和TPEF等非线性现象，提取有关线虫神经元结构和功能的有价值信息是可行的。
