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受激发射损耗荧光显微镜利用荧光饱和和激发态荧光受激损耗的非线性关系,通过限制损耗区域,可突破远场光学显微术的衍射极限分辨力并实现三维成像。基于对粒子速率方程组的修正,建立了描述荧光团各能级粒子数概率时间特性的模型,并定义了时间平均损耗效率判据。采用高斯函数模拟两束入射激光脉冲通过对模型的数值计算,模拟了激发脉冲的SIED激光脉冲的光强、脉冲宽度以及两束光的延迟时间等参量与损耗效率之间的关系,并获得了各参量的最佳值,优化了损耗效率,为提高系统分辨力提供了有效的途径。
The stimulated emission loss fluorescence microscope utilizes the nonlinear relationship between fluorescence saturation and excited state fluorescence excitation loss, and can overcome the diffraction limit resolution of far-field optical microscopy and achieve three-dimensional imaging by limiting the loss region. Based on the correction of the particle velocity equations, a model describing the temporal characteristics of the probability of each level of the fluorophore is established, and the criterion of time average loss efficiency is defined. Gaussian function was used to simulate the two laser pulses. The numerical simulation of the model, the relationship between the laser intensity and the pulse width of the SIED laser pulse and the delay time of the two laser beams were simulated and the loss efficiency was obtained. The optimal value of each parameter optimizes the loss efficiency and provides an effective way to improve the system resolution.