基于CCD虚拟针孔探测的双轴共焦显微技术

Virtual pinhole dual-axes confocal microscopy based on CCD detector

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。

In comparison with the conventional confocal microscopy,dual-axes confocal microscopy can use a low NA objective to achieve a high resolution,a large work distance and high S/N,due to the special dual-axes structure.The spatial resolution of a dual-axes confocal microscopy with virtual pinhole CCD is analyzed in theory and its corresponding experimental setup is established,and the axial response experiments are done using the established experimental setup.The full width at half a maxim（FWHM）in the axial direction is 2.63μm achieved by using an illuminating lens of NAi=0.117,a collecting lens of NAc=0.106 and θ=45°.And it is about 20 times higher than the axial FWHM of conventional confocal microscopy with the same parameters.