单光源双光路激光并行共焦测量系统设计

针对传统激光并行共焦测量过程中存在的泰伯效应,提出将数字微镜器件（DMD）引入激光并行共焦测量系统来正确辨识正焦面的位置。采用了DMD作为光分束器件,从理论上验证了它是一种投影式的阵列光源,对激光分束后不会在光路方向上产生泰伯像;同时,考虑DMD不能对分束后的光线产生会聚作用,并非高效的并行光源分束器件,本文将DMD与微透镜阵列(MLA)结合构建了单光源双光路并行共焦测量系统。该系统利用DMD光路探测正焦面位置,利用微透镜阵列光路进行精确的共焦测量。实验结果表明,两种光路下的正焦面位置仅相差2 μm,在一个泰伯间距范围之内,可以较好地克服泰伯效应对激光并行共焦测量的影响,进而保证较高精度的并行共焦测量。     

激光共焦透镜曲率半径测量系统

基于共焦技术独特的轴向层析定焦能力并结合气浮导轨平移台和激光干涉仪测长系统,研制了一套高精度、非接触激光共焦透镜曲率半径测量系统。该系统利用共焦轴向光强响应曲线的峰值点对应系统物镜聚焦焦点这一特性,使用峰值点对被测透镜的猫眼位置及共焦位置进行精确定位,并结合激光干涉仪获得透镜猫眼位置及共焦位置坐标值,从而计算得到透镜的曲率半径。系统由主控软件控制气浮导轨带动被测透镜在猫眼位置及共焦位置附近进行扫描测量,并实现信号采集和数据处理。实验表明,利用该系统测量透镜的曲率半径时,测量重复性优于2 μm,满足国内高精度透镜曲率半径测量的精度需求。该系统测量速度快、操作简便、结构简单且易于实现小型化。     

激光差动共焦透镜中心厚度测量系统的研制

基于高精度光学共焦定位技术研制了一种全新的非接触透镜中心厚度测量系统,该系统利用差动共焦技术的高轴向层析特性和轴向响应曲线的绝对零点对被测透镜的前表面顶点和后表面顶点分别进行精密瞄准定位;同时,利用激光干涉仪获得透镜前、后表面顶点的位置坐标;然后通过光线追迹算法计算透镜中心厚度,进而实现了透镜中心厚度的高精度非接触测量。实验结果表明,该系统测量精度高,测量标准差小于1μm,满足透镜中心厚度测量的精度要求。     

激光共焦高速扫描显微成像的高帧速重构算法

针对激光共焦扫描显微镜的往复式逐行扫描成像方式带来的帧图像数据分割难的问题,在分析系统扫描方式、振镜的实际运动方式与理论运动方式差异的基础上,利用相邻两帧图像相似性大的特点,提出了一套完整的高帧速重构算法。该算法通过连续帧特征区域差分的方式实现了一维信号序列的自适应分割,即实现了对一维信号序列进行动态排列及分割成二维阵列图像数据,从而重构出多帧高精度图像。实验表明,该算法的成像误差低于1.6%,适用于成像速度高达300帧/s的激光共焦扫描显微成像。     

激光共焦显微光束的偏转扫描

为了提高激光共聚焦系统的扫描速度,本文提出一种逐场扫描的场同步扫描方法。构建了激光共焦显微系统,将美国THORLABS公司的GVS002型二维检流计振镜应用于该系统,根据光学系统参数以及扫描范围要求计算振镜的整场扫描波形。借助NI公司的PCIe6353多功能数据采集卡,输出行同步的扫描波形,同时,对共焦显微系统共焦位置上针孔处的光强信号进行采集,先后扫描一幅256×256和512×512的图像,记录扫描图像和成像时间;然后,在相同的硬件结构下,以场同步的方式输出扫描波形,记录扫描图像和成像时间。实验结果表明:场同步方式扫描256×256图像的速度可提高10倍,扫描512×512图像的速度可提高5倍,且满足共焦显微成像的清晰、抗干扰能力强等要求。与行同步扫描方法相比,场同步扫描方法可以消除行与行之间转换的停留时间,在不改变硬件的情况下大幅提高扫描速度。     

基于共焦显微条件下线结构光共焦显微术的研究

美国的Minsky在20世纪50年代末提出共焦显微术的概念以来,尽管共焦显微术可以获得更高的分辨率,但不仅成像速度慢,而且需要使用光电增强器对采集光点信号进行增强,导致制造成本过高,所以除一些成像质量要求极高的显微系统外,其他的显微系统很少应用。讨论了线结构光共焦显微术的原理、优缺点以及影响性能的几个重要因素,并介绍其最新的研究状况。     

一种并行共焦显微镜的设计与研制

并行共焦显微镜是一种综合了光学、机械、电子、计算机及数字图像处理等技术,具有较高的横向和纵向分辨力的新颖三维表面无损检测仪器.为了保证其装配调节方便,在关键部件的结构设计上采用了微调结构,主要介绍了并行共焦显微镜的光学系统、物镜组设计、关键部件的微调结构设计及结构组成。     

激光差动共焦曲率半径测量系统的研制

针对国内高精度曲率半径计量需求,研制一套激光差动共焦曲率半径测量系统.该系统采用差动共焦定焦技术,利用轴向光强响应曲线的过零点精确对应物镜聚焦焦点这一特性,借助过零点对被测件的猫眼和共焦位置进行精密瞄准定位,通过干涉测长技术获取两点间的距离,继而实现曲率半径的高精度测量.该测量系统的机电控制由主控软件完成,可实现机电扫描、数据采集及数据处理,自动化程度高.实验证明,该系统定焦灵敏度高,受环境波动影响小,测量精度可达3×10-6,满足了高精度曲率半径的计量需求.     

曲面复眼成像系统的研究

研究了两种曲面复眼成像系统,并首次将曲面场镜阵列引入曲面复眼成像系统,使其边缘视场的成像质量进一步提高,视场角进一步加大。进行了成像系统的建模以及光线追迹,两种结构的视场角分别达到60°和88°,整个系统的体积分别为0.9 mm×0.9 mm×0.5 mm和0.9 mm×0.9 mm×0.75 mm。文中给出了用激光直写设备在曲面基底上进行光刻来制作曲面微透镜阵列的方法。     

基于数字微镜的共焦显微系统的光路设计

详细叙述了共焦技术中的横向扫描技术,介绍了基于数字微镜(DMD)的共焦显微镜结构与原理,建立了基于DMD的并行检测系统,并且进行了光路的优化设计.实验结果表明,采用传统共焦显微镜光路时,光线出射分光棱镜时存在棱镜内表面反射问题,导致DMD上同一像素块在CCD上成两个像.在此分析了上述现象产生的原理,给出了解决此问题的方...     

激光扫描共聚焦光谱成像系统

在激光扫描共聚焦显微成像技术基础上引入了光谱成像技术以便区分生物组织中的不同荧光成分。采用分光棱镜对荧光进行光谱展开,在光谱谱面处设置两个可移动缝片形成出射狭缝,两个步进电机带动安装其上的两个缝片设置系统在整个工作波长（400~700 nm）内的光谱带宽,其最小光谱带宽优于5 nm。用488 nm激光和低压汞灯实际测量了几条谱线对应的狭缝位置并和理论值做了比较,结果显示实际狭缝位置和理论值的差值均小于0.1 mm。在全光谱和50 μm出射狭缝（对应2.5 nm光谱带宽）对老鼠肾脏组织进行了共聚焦光谱成像实验,获得了老鼠肾脏组织中DAPI标定的细胞核图像和Alexa Fluor^®488标定的肾脏小球曲管图像,实现了对老鼠肾脏组织不同成分的区分。实验结果表明：提出的系统能够进行共聚焦光谱成像,扩大了共聚焦显微镜的适用范围。     

基于并行共聚焦显微系统的物方差动轴向测量

差动共聚焦显微成像技术可以获得很高的轴向测量精度,然而已有的差动共聚焦测量技术主要适用于激光扫描共聚焦,还不能满足微纳加工过程中对工件进行非接触式的在线、在位测量的要求。本文在分析差动共聚焦显微成像系统能够实现轴向测量原理的基础上,提出了适用于并行共聚焦技术的轴向测量方法。该方法利用均匀白光照明,在像方只需要使用一台相机做探测器,在物方通过移动载物台分别对样品在焦前和焦后两次成像,根据预先刻度好的差动曲线就可以得出物体表面的高度。理论模拟与实验结果均表明,该方法可以实现高精度的轴向测量,对500nm的台阶样品测量的平均误差为2.9nm,相对误差为0.58%。该方法简单、廉价、测量精度高,可以用于普通显微镜,易于实现样品的三维快速形貌还原与测量。     

Radon track imaging in CR-39 plastic detectors using confocal scanning laser microscopy

Confocal scanning laser microscopy (CSLM) has been used to provide the first images of radon track populations in two external CR-39 plastic detectors. Measurements of variables including track area distribution and estimates of the angle of track inclination (dip) derived from surface CSLM sections are presented. CSLM depth slices, combined with three-dimensional (3D) visualization techniques, provide a new, non-destructive way of examining the 2D and 3D geometry of the etched tracks within solid-state nuclear track detectors that may prove useful in complementing existing optical microscopy methods.     

Double-pulse fluorescence lifetime imaging in confocal microscopy

A theoretical analysis of a new technique for fluorescence lifetime measurement, relying on (near steady state) excitation with short optical pulses, is presented. Application of the technique to confocal microscopy enables local determination of the fluorescence lifetime, which is a parameter sensitive to the local environment of fluorescent probe molecules in biological samples. The novel technique provides high time resolution, since it relies on the laser pulse duration, rather than on electronic gating techniques, and permits, in combination with bilateral confocal microscopy and the use of a (cooled) CCD, sensitive signal detection over a large dynamic range. The principle of the technique is discussed within a theoretical framework. The sensitivity of the technique is analysed, taking into account: photodegradation, the effect of the laser repetition rate and the effect of non-steady-state excitation. The features of the technique are compared to more conventional methods for fluorescence lifetime determination.     

Three‐dimensional imaging of porous media using confocal laser scanning microscopy

In the last decade, imaging techniques capable of reconstructing three‐dimensional (3‐D) pore‐scale model have played a pivotal role in the study of fluid flow through complex porous media. In this study, we present advances in the application of confocal laser scanning microscopy (CLSM) to image, reconstruct and characterize complex porous geological materials with hydrocarbon reservoir and CO₂ storage potential. CLSM has a unique capability of producing 3‐D thin optical sections of a material, with a wide field of view and submicron resolution in the lateral and axial planes. However, CLSM is limited in the depth (z‐dimension) that can be imaged in porous materials. In this study, we introduce a ‘grind and slice’ technique to overcome this limitation. We discuss the practical and technical aspects of the confocal imaging technique with application to complex rock samples including Mt. Gambier and Ketton carbonates. We then describe the complete workflow of image processing to filtering and segmenting the raw 3‐D confocal volumetric data into pores and grains. Finally, we use the resulting 3‐D pore‐scale binarized confocal data obtained to quantitatively determine petrophysical pore‐scale properties such as total porosity, macro‐ and microporosity and single‐phase permeability using lattice Boltzmann (LB) simulations, validated by experiments.     

激光共聚焦扫描显微镜的光学设计

为获得较高分辨率的细胞图像,设计了激光共聚焦光学系统。通过较复杂结构物镜实现了照明光路系统和发射光路系统的设计。用Zemax对照明光路和发射光路进行了设计,仿真过程中照明光路的聚焦弥散斑直径小于1μm,照明针孔处的聚焦弥散斑直径小于20μm,发射光路的聚焦弥散斑直径小于20μm,同时照明光路和发射光路的MTF曲线接近衍射极限,达到较理想的情况。     

In-vivo confocal scanning laser microscopy of the cerebral microcirculation

Confocal scanning laser microscopy (CSLM) was used to study the microcirculation of the brain neocortex in anaesthetized rats. After removal of the dura mater, implantation of a closed cranial window, and intravenous injection of fluorescein, three-dimensional reconstructions of cortical capillaries were performed down to a depth of 250 μm below the pial surface. Using a one-dimensional approach (single line scanning), erythrocyte (negative contrast in fluorescently labelled plasma) and leucocyte (labelled with rhodamine 6 G) velocity and supply rate in cortical capillaries were measured. The effect of CO₂-inhalation on capillary blood flow dynamics was studied. Capillaries were imaged continuously for up to 1 h without changes in flow or fluorescence pattern. However, by increasing the laser power 10–100-fold, aggregate formation was induced and capillaries were occluded, possibly due to damage to vascular endothelium. We conclude that CSLM can be used to study morphological and dynamic aspects of fluorescently labelled subsurface structures in organs of experimental animals.     

Three-dimensional imaging in fluorescence by confocal scanning microscopy

The improved resolution and sectioning capability of a confocal microscope make it an ideal instrument for extracting three-dimensional information especially from extended biological specimens. The imaging properties, also with finite detection pinholes are considered and a number of biological applications demonstrated.