基于微透镜阵列和振镜扫描的光谱分辨多焦点多光子显微技术

Spectrally Resolved Multifocal Multiphoton Microscopy Using Microlens Array and Galvo Mirror Scanning

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。

We present a novel spectrum-resolved multifocal multiphoton microscopy（SR-MMM） that is capable of performing fast 2-dimensional（2D） spectral measurements of fluorescent samples with optical sectioning.Fluorescence emission from the excited lines on the sample is spectrally dispersed with a prism,and a CCD array is used to acquire the spectrally resolved image.Spectrally resolved images of 512 pixel×512 pixel can be obtained by acquiring only 128 raw images when a 4×4 excitation foci array is used.Multi-color fluoresent beads,sample slide of a stained lily and pollen particles are used for spectrally resolved imaging experiment.The spectral measurement range is from 450 nm to 700 nm,and the spectral resolution is less than 3 nm.