An optical technique for remote focusing in microscopy

We describe the theory of a new method of optical refocusing that is particularly relevant for confocal and multiphoton microscopy systems. This method avoids the spherical aberration that is common to other optical refocusing systems. We show that aberration-free refocusing can be achieved over an axial scan range of 70 μm for a 1.4 NA objective lens. As refocusing is implemented remotely from the specimen, this method enables high axial scan speeds without mechanical interference between the objective lens and the specimen.     

基于偏振衍射色散共焦的光学元件轴向间距测量

光学元件轴向间距测量对精密光学系统的定位和装调具有重要意义.针对现有色散共焦测量系统中色散物镜结构复杂、色散范围小的问题,提出基于偏振衍射色散共焦的光学元件轴向间距测量方法,将传统折射共焦镜头几百毫米的轴向尺寸和复杂的装调需求简化为数毫米的单片镜,简化了系统结构.透镜间距和厚度测量实验表明,间距测量误差为10μm.     

Improvement of the axial trapping effect using azimuthally polarised trapping beam

A dual optical tweezers system,which consists of a doughnut mode optical tweezer (DMOT) with the azimuthally polarised trapping beam and a solid mode optical tweezer (SMOT) with the Gauss trapping beam was constructed to compare the axial trapping effect of DMOT and SMOT.The long-distance axial trapping of ST68 microbubbles (MBs) achieved by DMOT was more stable than that of SMOT.Moreover the axial trapping force measured using the viscous drag method,was depended on the diameter of the particle,the laser power,and the numerical aperture (NA) of the objective lens.The measurement of the axial trapping force and the acquisition of CCD images of trapping effect confirmed that the DMOT showed excellent axial trapping ability than SMOT.A simple and effective method is developed to improve axial trapping effect using the azimuthally polarized beam as trapping beam.This is helpful for the long-distance manipulating of particles especially polarised biological objects in axial direction.     

绿光光存储实验研究

在光存储技术中,采用短激光波长激光器和大数值孔径物镜是提高光存储密度的两种有效途径.本文采用532 nm波长激光, 0.65数值孔径物镜,建立了绿光存储实验装置.并采用该实验装置在CD-RW相变光盘上进行了存储实验研究,实现了线宽约为500 nm的实验结果.     

Depth of focus extended microscope configuration for imaging of incorporated groups of molecules, DNA constructs and clusters inside bacterial cells

Imaging of small objects such as single molecules, DNA clusters and single bacterial cells is problematic not only due to the lateral resolution that is obtainable in currently existing microscopy but also, and as much fundamentally limiting, due to the lack of sufficient axial depth of focus to have the full object focused simultaneously. Extension in depth of focus is helpful also for single molecule steady state FRET measurements. In this technique it is crucial to obtain data from many well focused molecules, which are often located in different axial depths.In this paper we present the implementation of an all-optical and a real time technique of extension in the depth of focus that may be incorporated in any high NA microscope system and to be used for the above mentioned applications. We demonstrate experimentally how after the integration of special optical element in high NA 100× objective lens of a single molecule imaging microscope system, the depth of focus is significantly improved while maintaining the same lateral resolution in imaging applications of incorporated groups of molecules, DNA constructs and clusters inside bacterial cells.     

光谱共焦三维形貌仪用超大色散线性物镜组设计

光谱共焦三维形貌仪具有精度高、测量范围大、适应性强等特点,其关键部件是产生轴向色散的物镜,色散物镜轴向色散与波长间的非线性会导致系统整体性能下降。分析了共焦法检测表面形貌的工作原理,以及产生线性轴向色散的条件,根据共焦光学系统的基本设计原理,在预设的设计指标及要求下,完成一个超大色散线性物镜组设计。该色散物镜组采用四级串联结构,在400~700 nm光谱范围内达到30 mm的色散范围,波长离焦量和波长之间线性度通过线性回归拟合得出判定系数为1,满足色散的线性度要求。     

照相物镜移植应用的研究

对于用逐步修改法设计光学系统,初始结构的选择是重要环节。除在典型形式中寻求初始结构外,也可以借鉴其它系统的光学结构进行相似创新。照相物镜移植应用指的是不同系统物镜因成像功能相似,故可实施光学结构的优化替代。本文研究了各种物镜成像功能的相似性,不同系统大孔径、大视场物镜光学特性的边缘性;根据像差理论阐明了照相物镜是大孔径、大视场系统理想的结构替代源;结合照相物镜丰富的光学结构资源的实际情况,指出移植应用是可行的。将照相物镜移植应用于其它系统大孔径、大视场物镜设计中,生产出与照相物镜结构相似、像质优良的镜头。实践证明了移植应用的实用性与可操作性。     

采用变焦液体透镜的共焦检测系统的设计与仿真

共焦显微技术越来越广泛地应用于微机电系统和半导体器件的三维轮廓测量。设计一种新型采用变焦透镜作为轴向扫描方式的共焦显微系统,利用变焦透镜的焦距变化来代替传统的轴向位移扫描。系统实现了无机械运动的轴向扫描,消除了传统共焦系统中由位移台移动带来的振动,不仅减小了系统的复杂度,而且降低了成本。进一步设计了基于位移台和变焦透镜的两种共焦检测系统,并且对其进行了仿真实验。实验结果表明:基于位移台和基于变焦透镜的共焦系统的聚焦成像和离焦成像结果类似,并且光强的轴向分布曲线也基本相同,表明此方法可行。     

Tomographic imaging of a suspending single live cell using optical tweezer-combined full-field optical coherence tomography

We propose a label-free depth-resolved tomographic scheme for imaging a single live cell in fluid. This approach utilizes a modified time-domain full-field optical coherence tomography (FF-OCT) system combined with an optical tweezer technique. The optical trap for holding a moving specimen is made by tightly focusing a 1064 nm Q-switching pulsed laser beam with a 1.0 NA microscope objective in the sample arm of the FF-OCT part. By cosharing the probe for both systems, the optical actions of trapping and cellular resolution tomographic imaging could be achieved simultaneously. Feasibility of the combined system is demonstrated by imaging micron-sized polystyrene beads and a living suspension cell in medium.     

基于相位复原技术测试高数值孔径光学成像系统

干涉测量的方法已被广泛应用于光学系统的波前测试中。但是由于高数值孔径的标准具加工成本高、风险大,使用干涉测量的方法对高数值孔径光学系统进行测试存在困难。运用相位复原技术对高数值以小孔衍射光束作为高精度测试基准波前进行孔径光学系统波前测试,可以解决上述问题。对不同频率波前形成的点扩散函数模拟,分析了实际测试所需的实验条件。由于缺乏高精度的对比实验条件,引入一种新的误差分析方法,搭建检测平台完成对显微镜光学系统的波前测试。通过验证实验,证明了该测试方法的可靠性。     

Demonstration of a no-moving-parts axial scanning confocal microscope using liquid crystal optics

For the first time, to the best of authors’ knowledge, a no-moving-parts axial scanning confocal microscope (ASCM) system is designed and demonstrated using a combination of a large diameter liquid crystal (LC) lens and a classical microscope objective lens. By electrically controlling the 5 mm diameter LC lens, the 633 nm wavelength focal spot is moved continuously over a 48 μm range with a measured 3-dB axial resolution of 3.1 μm using a 0.65 numerical aperture (NA) micro-objective lens. The ASCM is successfully used to image an Indium Phosphide (InP) twin square optical waveguide sample with a 10.2 μm waveguide pitch and 2.3 μm height and width. Using fine analog electrical control of the LC lens, a super-fine sub-wavelength axial resolution of 270 nm is demonstrated. The proposed ASCM can be useful in various precision three-dimensional (3D) imaging and profiling applications.     

Tight focusing of a double-ring-shaped, azimuthally polarized beam

We study the focusing properties of a double-ring-shaped azimuthally polarized beam through an annular high NA objective lens. It is shown that a subwavelength focal hole (～0.5λ) with a quite long depth of focus (～26λ) is achieved near the focus. This kind of nondiffracting focal hole is called dark channel, which may have applications in atom optical experiments, such as with atomic lenses, atom traps, and atom switches.     

基于CCD虚拟针孔探测的双轴共焦显微技术

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。     

Optical design of a full-pupil field,non-contact and high resolution corneal curved objective lens

It has been a challenge to overcome the corneal curvature radius to design a full-pupil field, non-contact and high resolution corneal curved objective lens, which covers the cornea full-pupil field and has the ability to resolve corneal cells. In this paper, we report an optical design of a full-pupil field, non-contact corneal curved objective lens for high resolution cornea imaging. The advantages of this lens are that it has a wide field of view (FOV) with the corneal curved image surface, maintains the beam normal incidence, as well as non-contact lens imaging, and offers a cell-level lateral resolution of cornea structure. The analysis of optimization shows that the system achieves diffraction limit in a circular FOV of 4 mm diameter covering the full-pupil zone. The theoretical lateral resolution is about 2.5 μm with an image space NA of 0.16, which is sufficient to resolve corneal cells of 7 μm diameter, and the working distance is larger than 15 mm which is enough for a non-contact objective lens. So the optical design is effectively and efficiently meeting the demand of specifications.     

Spring constant modulation in a zone plate tweezer using linear polarization

At large NAs a micro-Fresnel zone plate produces a focal spot that is more elliptical than that produced by an objective lens with the same NA. Using this anisotropy we demonstrate a method for modulating the spring constant of an optical trap by rotating the linear input polarization. The focal spot ellipticity is enhanced by the apodization factor of the zone plate and its extremely high NA. By measuring the positions of trapped particles we obtain two-dimensional histograms of particle position. These indicate that the trap spring constant is 2.75 times larger perpendicular to the incident polarization than along it. The elliptical focal spot distribution can be rotated by rotating the incident polarization, allowing the spring constant along a given direction to be modulated.     

Tightly focusing of spirally polarized Quadratic Bessel Gaussian beam through a dielectric interface

Based on vector diffraction theory, the tight focusing properties of spirally polarized axisymmetric Bessel-modulated Gaussian beam through a dielectric interface on high numerical aperture (NA) are investigated theoretically. The optical intensity distribution in the focal region of high NA objective lens is investigated in detail by numerical calculations. The results show that the focal shift induced in the focal region is by mismatch of refractive indices across the dielectric interface. It is also found that the optical intensity in focal region of spirally polarized Quadratic Bessel Gaussian (QBG) beam can be altered considerably by changing spiral parameter C that indicates the polarization spiral degree of the incident beam.     

高数值孔径聚焦三维光链的研究

通过设计衍射光学元件对入射矢量光进行调制，在高数值孔径聚焦系统焦点附近产生沿光轴方向的三维多点光俘获结构——光链.并针对不同的入射矢量偏振、聚焦透镜的数值孔径以及衍射光学元件结构，对光链性能的影响分别进行了系统的分析，实现对该独特光俘获结构的可控性研究. 关键词： 衍射光学元件 矢量光 光镊     

Influence on the resolution of target detection with defective NR-PC flat lens incorporating Al,Cu by dynamic scanning scheme

Based on the defective NR-PC flat lens incorporating Al or Cu, the effects on the resolution for target detection and imaging are studied with dynamic scanning scheme. We use the finite-difference-time-domain method to do this research. Firstly, by using the NR-PC flat lens may improve the refocusing resolution by four times, if compared to the directly backscattered lightwave without using the NR-PC lens. Then, incorporating Al or Cu into the NR-PC flat lens, we find that the dope modes with very high quality factors will occur in the photonic forbidden band. Also the stimulated radiation is enhanced for the character of energy localization of defective photonic crystal. Further studies show that different kinds of metal and structures will result in different dopped modes in the photonic forbidden band, which will lead to the change of the corresponding resolution. Due to the displacement field redistribution, the photonic crystal containing metal ingredients may take on the typical qualities of all-directional reflection, and the resonant tunneling effect may effectively reduce the loss, which provides greatly improvement on the refocusing resolution when using the defective NR-PC flat lens. In conclusion, our investigation provided the basis for converting an idealized LHM lens into a physically realizable NR-PC flat lens. Meanwhile, by impregnating metallic defect into the NR-PC lens, a new method is found out for the improvement of the refocusing resolution, which further optimizes the performance of a detecting and imaging system.     

Characterization of aplanatic hologram lenses for optical disk pick-ups

Imaging characteristics for holographic objective lenses application to optical disk pick-ups was analyzed by a comparison between flat and spherical aplanatic hologram lenses with the aid of the third order aberration theory and ray tracing method. The aplanatic hologram lens image field was 0.5 deg for NA=0.5 with focal length f=5 mm under the Marechal criterion. The results indicates that the aplanatic hologram lenses are feasible for optical disk applications.     

有限远共轭显微镜光镊设计和误差分析

光镊是研究单分子生物物理特性的独特工具, 因而光镊设备的研发是一个极为重要的课题. 本文根据矩阵光学, 对基于有限远共轭显微镜的光镊操控光路进行计算, 得出了阱位径向操控和轴向操控方程, 并分析了光束调控系统、 共焦系统后置透镜和耦合透镜安装位置误差及物镜轴向位置调整对光镊阱位径向及轴向操控精度的影响. 计算结果显示, 当物镜初级像面和耦合透镜像方焦面完全重合, 光束调控系统和耦合透镜的距离误差对阱位径向和轴向操控精度没有影响. 光镊系统元器件定位不准时, 基于无限远共轭显微镜光镊的阱位径向操控误差和轴向操控误差都小于基于有限远共轭显微镜光镊的阱位径向操控误差和轴向操控误差. 当光镊耦合透镜定位误差控制在小于10 mm时, 基于有限远共轭显微镜光镊的径向和轴向操控误差分别小于5.9%和11.4%, 有限远共轭显微镜仍然存在改造为光镊的价值.本文理论为基于有限远共轭显微镜的光镊设计、改造和操控提供理论和实验指导. 关键词： 光镊 光学设计 矩阵 误差