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双轴共焦显微技术具有独特的非共轴结构,与传统单轴共焦显微技术相比可利用较低数值孔径物镜实现较高的轴向分辨力,且具有工作距离大、信噪比高等优势。对基于CCD虚拟针孔(VPH)探测的双轴共焦显微成像系统的空间分辨特性进行了理论分析,并构建了相应的实验系统,对其轴向响应进行了实验验证。实验中照明物镜NAi=0.117,采集物镜NAc=0.106,θ=45°,得出系统轴向半高宽(FWHM)为2.63μm,比同等参数(NA=0.117)下单轴共焦显微系统的轴向FWHM高出约20倍。
Biaxial confocal microscopy has a unique non-coaxial structure, compared with the traditional uniaxial confocal microscopy can use a lower numerical aperture objective lens to achieve higher axial resolution, and has a large working distance, high signal-to-noise ratio advantages. The spatial resolution characteristics of biaxial confocal microscopy imaging system based on virtual pinhole (VPH) detection of CCD are theoretically analyzed and a corresponding experimental system is constructed. The axial response is verified by experiments. In the experiment, NAi = 0.117, NAc = 0.106 and θ = 45 ° were collected. The FWHM of the system was 2.63μm, which was higher than that of the uniaxial confocal microscope system with the same parameter (NA = 0.117) About 20 times higher than FWHM.