多焦点多光子显微(MMM)和单点扫描多光子显微相比,可显著提高光能利用率和成像速度,在生命科学研究领域具有广泛的应用前景。但现有MMM技术成像质量和成像速度之间存在矛盾,增加单位面积内的焦点数目可提高成像速度,但过小的焦点间距离会产生子光束串扰,从而影响纵向空间分辨率。利用Zemax软件模拟产生了高密度激发点阵,将传统的不小于6 mm的相邻焦点间距缩小到约3 mm;分析了高密度点阵激发成像产生噪声的原因;根据时间复用技术的原理设计正方形时间延板,对相邻子光束脉冲进行不同时间的延迟,从而可以消除MMM系统中高密度激发点阵子光束间的串扰。 Multi-focal multi-photon microscopy (MMM) can significantly improve the utilization of light energy and imaging speed compared with single-point scanning multiphoton microscopy and has wide application prospect in the field of life science research. However, there is a conflict between the imaging quality and the imaging speed of the existing MMM technology. Increasing the number of focal points per unit area can increase the imaging speed, but the too small inter-focus distance generates sub-beam crosstalk, which affects the longitudinal spatial resolution. The Zemax software was used to simulate the high-density excitation lattice. The traditional focus distance of less than 6 mm was reduced to about 3 mm. The reason of high-density lattice imaging was analyzed. According to the principle of time multiplexing The square time delay board is designed to delay adjacent sub-beam pulses at different times, so as to eliminate the crosstalk between the high-density excitation sub-beam of the MMM system.