分析了多路频分复用并行显微荧光探测的基本原理,即把激发光分成多束,对每一束进行不同频率调制,聚焦到生物样品的不同位置,激发产生相应频率的荧光信号,再对光电倍增管(PMT)接收的荧光信号进行分频解调,实现实时、并行和高分辨率的探测技术。搭建了激发光源为405 nm近紫外激光的双路方波调制荧光显微探测系统,实验探测了老鼠神经细胞显微形态,分析解调了双点荧光能量随时间变化的曲线。并研究了荧光显微成像系统放大率、时间分辨率等技术参数,还通过数值分析给出了避免各通道间信号串扰的条件。 The basic principle of multi-channel frequency-division multiplexing parallel micro-fluorescence detection is analyzed. That is, the excitation light is divided into multiple beams, each of the beams is modulated at different frequencies, focused on different positions of the biological sample, and excited to generate fluorescence signals of corresponding frequencies, Then the PMT received fluorescence signal frequency division demodulation, real-time, parallel and high-resolution detection technology. A two-way square-wave fluorescence microscopy system with excitation light source of 405 nm near-UV laser was set up. The microscopic morphology of neuronal cells in mice was probed and the curve of energy of two-point fluorescence with time was analyzed and demodulated. The technical parameters such as magnification and time resolution of fluorescence microscopy imaging system are also studied. The condition of avoiding crosstalk between channels is given by numerical analysis.