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提出一种具有快速层析成像以及光谱分辨功能的多光子激发荧光显微技术。采用微透镜阵列产生激发光点阵,利用线扫描方式扫描阵列点,对样品进行多线并行多光子激发,利用棱镜色散荧光信号,同时,利用面阵CCD并行记录光谱分辨的多线荧光信号。采用4×4的微透镜阵列,仅需要记录128幅图像,即可重构512 pixel×512 pixel的光谱分辨荧光显微图像。对多色荧光珠、染色铃兰根茎以及花粉颗粒等样品进行实验,得到样品的双光子激发荧光光谱分辨图像,光谱测量范围为450~700 nm,光谱分辨率为3 nm。
A multi-photon excited fluorescence microscopy with fast tomography and spectral resolution is proposed. The microlens array was used to generate the array of excitation light. The array was scanned by line scanning. The sample was multi-line parallel multiphoton excitation. The prism was used to disperse the fluorescence signal. Meanwhile, the parallel multi-line fluorescence signal was recorded by the array CCD. With a 4 × 4 microlens array, only 128 images need to be recorded to reconstruct a 512 pixel × 512 pixel spectrally resolved fluorescence microscopy image. Two-photon excitation fluorescence spectra of the samples were obtained by experiments on samples of multicolored fluorescent beads, stained lily of the valley and pollen grains. The measurement range was 450-700 nm and the spectral resolution was 3 nm.