系统畸变对共焦显微成像分辨率的影响
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摘要
介绍了一种基于径向偏振光束和光瞳滤波的超分辨共焦显微成像系统,并基于矢量和标量衍射理论,研究了系统畸变,包括探测针孔尺寸及针孔偏移等对成像分辨率的影响。介绍了系统结构及工作原理,并推导了针孔大小变化及发生偏移时,成像系统强度响应函数的数学表达式;然后,通过设置不同的针孔尺寸大小及针孔偏移量进行了数值计算,结果表明,当针孔归一化半径分别为(和10(时,系统强度响应的横向半高宽分别为0.488和0.492,轴向半高宽分别5.1和9.1。假设探测器与针孔之间没有相对位移,针孔的横向偏移会对系统的轴向分辨率产生影响。针孔的轴向偏移不会对系统的分辨率产生影响,但是会使探测器接收到的响应强度产生相移,相移量由针孔偏移大小来决定。研究结果对于超分辨共焦显微成像技术的发展及应用提供了一定的指导意见。
In this paper,a super-resolution confocal microscopy system based on radial polarized beam and pupil filtering is introduced,and the system distortion is studied based on the vector and scalar diffraction theories,including the effect of pinhole size and pinhole displacement on imaging resolution. This paper introduces the structure and working principle of the system,and deduces the mathematical expression of the intensity response function of the imaging system when the pinhole size changes and the offset occurs. Then,through the numerical calculation for different pinhole sizes and pinhole offsets,the results show that when the normalized radius of pinhole are( and 10( respectively,the transverse half-widths with half maximum( HWHM) of the system strength response are 0. 488 and 0. 492 respectively,and the axial HWHMs are 5. 1 and 9. 1 respectively. When there is no relative displacement between the detector and the pinhole,the transverse displacement of the pinhole will have an effect on the system's axial resolution. The axial displacement of the pinhole will not affect the resolution of the system,but it will cause the response strength of the detector to shift and the shift is determined by the displacement of the pinhole. The research results provide guidance for the development and applications of the super-resolution confocal microscopy.
In this paper,a super-resolution confocal microscopy system based on radial polarized beam and pupil filtering is introduced,and the system distortion is studied based on the vector and scalar diffraction theories,including the effect of pinhole size and pinhole displacement on imaging resolution. This paper introduces the structure and working principle of the system,and deduces the mathematical expression of the intensity response function of the imaging system when the pinhole size changes and the offset occurs. Then,through the numerical calculation for different pinhole sizes and pinhole offsets,the results show that when the normalized radius of pinhole are( and 10( respectively,the transverse half-widths with half maximum( HWHM) of the system strength response are 0. 488 and 0. 492 respectively,and the axial HWHMs are 5. 1 and 9. 1 respectively. When there is no relative displacement between the detector and the pinhole,the transverse displacement of the pinhole will have an effect on the system's axial resolution. The axial displacement of the pinhole will not affect the resolution of the system,but it will cause the response strength of the detector to shift and the shift is determined by the displacement of the pinhole. The research results provide guidance for the development and applications of the super-resolution confocal microscopy.
引文
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[4]刘嘉兴,周哲海,祝连庆,刘超,郭阳宽.超分辨显微成像环形光瞳的结构设计[J].激光杂志,2016,37(12):5-8.
[5]赵维谦,唐芳,邱丽荣,刘大礼.轴对称矢量光束聚焦特性研究现状及其应用[J].物理学报2013,62(5):1-7.
[6]阎杰,鲁拥华,王沛明海.径向偏振光聚焦光斑研究[J].光学学报,2012,30(12):3598-3602.
[7]GE Huayong,WANG Baohua,REN Qiushi.Influence of Detector on Resolution in Confocal Imaging System[J].光子学报,2006,35(4):587-589.
[8]周哲海.轴对称偏振光束的生成、特性及应用[J].北京:清华大学,2010.
[2]WILSON T and Sheppard C J R.Theory and Practice of Scanning Optical Microscopy[M].London:Academic Press,1984.
[3]ZHAO W Q,TAN J B,QIU L R.Tri-heterodyne confocal microscope with axial superresolution and higher SNR[J].Opt.Exp.12(21):5191~5197(2004).
[4]刘嘉兴,周哲海,祝连庆,刘超,郭阳宽.超分辨显微成像环形光瞳的结构设计[J].激光杂志,2016,37(12):5-8.
[5]赵维谦,唐芳,邱丽荣,刘大礼.轴对称矢量光束聚焦特性研究现状及其应用[J].物理学报2013,62(5):1-7.
[6]阎杰,鲁拥华,王沛明海.径向偏振光聚焦光斑研究[J].光学学报,2012,30(12):3598-3602.
[7]GE Huayong,WANG Baohua,REN Qiushi.Influence of Detector on Resolution in Confocal Imaging System[J].光子学报,2006,35(4):587-589.
[8]周哲海.轴对称偏振光束的生成、特性及应用[J].北京:清华大学,2010.