双光子荧光与相干反斯托克斯拉曼散射同属于三阶非线性效应,二者之间的差异与联系是一个值得研究的问题.本文基于自行搭建的超连续谱近红外宽带相干反斯托克斯拉曼散射显微成像系统进行光谱成像,同时通过理论与实验对比分析了双光子荧光与相干反斯托克斯拉曼散射图像存在差异的原因.结果表明,具有亚微米以上横向分辨率的相干反斯托克斯拉曼散射成像系统,可以使用较大尺寸的荧光珠进行双光子荧光成像,通过解卷积得到双光子荧光成像的系统分辨率,并将它近似等效于相干反斯托克斯拉曼散射成像系统的当下分辨率.如果需要得到相干反斯托克斯拉曼散射成像系准确的分辨率结果,就必须使用尺寸比相干反斯托克斯拉曼散射成像系统实际分辨率小的球形样品进行实验测量.
Two-photon excitation fluorescence (2PEF) and coherent anti-Stokes Raman scattering (CARS) are both third-order nonlinear optical processes, but for a long time, the true relationship and differences between them are not clearly understood. For decades, the second harmonic generation has been studied in conjunction with two-photon excitation fluorescence, so it was thought that the latter was a second-order nonlinear optical process. In order to make the two nonlinear interaction processes clear enough, the two nonlinear interaction processes are worthy to study at the same time. In this paper, firstly, we give the relationships between the 2PEF, CARS signal and their third-order nonlinear susceptibility, respectively; secondly, we use our own near infrared super-continuum CARS microscopy system to study both processes. In doing so, we describe the relationship between their third-order nonlinear susceptibility and the signal. The reconstructed images derived from CARS and those derived from 2PEF differ significantly when imaging the same 1.01 $/muup$m fluorescence polystyrene beads. If the lateral spatial resolution of the CARS imaging system is larger than the fluorescence polystyrene beads, the measured size cannot be used to calculate the real spatial resolution of the CARS system. However, the resolution of the 2PEF microscopy system can be obtained through the de-convolution of the 2PEF image, which is approximately equivalent to the current resolution of the CARS imaging system, which is measured using 280 nm polystyrene beads. The images of 280 nm polystyrene beads and 190 nm fluorescent polystyrene beads also exhibit differences between the two samples and the environment around them, respectively. This means that although CARS and 2PEF are both third-order nonlinear optical processes, they have their own properties. In particular, CARS is a third-order nonlinear optical oscillation process which is caused by the phasing match condition, but 2PEF is not influenced by the phasing match condition