激光扫描共聚焦光谱系统设计

Design of laser scanning confocal spectroscopy system

为提高激光扫描共聚焦显微镜光谱模块的分光性能,设计了双Amici棱镜光谱结构。根据棱镜色散特性,计算得到Amici棱镜和双Amici棱镜的线性与非线性数学模型,给出直视型棱镜结构参数。比较相同色散条件下Amici和双Amici棱镜结构,得到双Amici棱镜组能提供更好的色散线性度。在光谱波段400~700 nm,中心波长偏向角为0°,最大色散角为4°,光谱分辨率为5 nm的条件下,利用双Amici棱镜结构设计共光轴直视型分光系统。用双Amici棱镜结构和单棱镜对比,进行了光谱分布实验。实验结果表明,双Amici棱镜组的色散线性度明显优于单棱镜,大大改善了因棱镜非线性分光特性对荧光成像光谱分辨率的影响。同时,将双Amici棱镜结构设计进共聚焦光谱成像模块中,对牛肺动脉内皮细胞切片进行了三通道荧光成像,得到了高质量的多通道荧光图像。

A spectral structure for the double Amici prism was designed to improve the spectroscopic performance of a laser scanning confocal microscope spectroscopy module.Linear and nonlinear mathematical models were formulated for the Amici and double Amici prisms,and the structure parameters of directviewing prism were chosen according to the prism dispersion characteristics.The structures of the Amici and double Amici prisms were compared for the same dispersion conditions,and the double Amici prism group was found to provide better dispersion linearity.A common optical axis direct vision spectroscopic system was developed using a double Amici prism for a 400-700 nm spectral range and 5 nm spectral resolution,with central wavelength deviation angle of 0°and maximum dispersion angle of 4°.The experimental results show that the dispersion linearity of this structure is significantly better than that of a single prism.Spectral resolution issues caused by the nonlinear spectroscopic characteristics of prisms are greatly reduced.