Tai Shih-Peng, Chan Ming-Che, Tsai Tsung-Han, Guol Shi-Hao, Chen Li-Jin, Sun Chi-Kuang
Opt Express. 2004 Dec 13;12(25):6122-8. doi: 10.1364/opex.12.006122.
Self-phase-modulation and group velocity dispersion of near IR femtosecond pulses in fibers restrict their use in two-photon fluorescence microscopy (TPFM). Here we demonstrate a hollow-core photonic crystal fiber based two-photon fluorescence microscope with low nonlinearity and dispersion effects. We use this fiber-based TPFM system to take two-photon fluorescence (chlorophyll) images of mesophyll tissue in the leaf of Rhaphidophora aurea. With less than 2mW average power exposure on the leaf at 755nm, the near zero-dispersion wavelength, chloroplasts distribution inside the mesophyll cells can be identified with a sub-micron spatial resolution. The acquired image quality is comparable to that acquired by the conventional fiber-free TPFM system, due to the negligible temporal pulse broadening effect.
近红外飞秒脉冲在光纤中的自相位调制和群速度色散限制了它们在双光子荧光显微镜（TPFM）中的应用。在此，我们展示了一种基于空心光子晶体光纤的双光子荧光显微镜，其具有低非线性和色散效应。我们使用这个基于光纤的TPFM系统获取了金绿宝石叶肉组织的双光子荧光（叶绿素）图像。在755nm（近零色散波长）下，对叶片的平均功率曝光低于2mW时，叶肉细胞内叶绿体的分布能够以亚微米空间分辨率被识别。由于时间脉冲展宽效应可忽略不计，所获取的图像质量与传统无光纤TPFM系统获取的图像质量相当。
