结构光三维成像系统的计算机仿真

基于相位测量轮廓术和摄像机模型,提出一种结构照明三维成像系统的高精度计算机仿真算法。对于给定的物体三维模型,首先根据系统结构,采用Z缓冲技术消除遮挡和阴影部分,得到与摄像机像素点对应的物体表面采样点三维坐标,再使用统一的数学模型和方法处理投影过程。根据摄像机及投影仪的内、外参数,最终得到了摄像机像素点、物体表面采样点和投影仪像素坐标三者之间的对应关系,从而实现了结构照明三维成像系统仿真。为实际系统的结构设计、调整和参数校正提供了参考。     

隐失驻波全内反射荧光显微术的数值计算

从理论上详细计算了在全内反射的条件下，两束入射光产生的隐失干涉场(即隐失驻波)的强度分布，并分析了其不同于传统传播波干涉场的特点。同时使用数值模拟证明了利用隐失干涉场，即隐失驻波的激发方式，可以提高系统的分辨力，在横向实现超经典分辨的荧光成像。具体的分析表明，两束光以相等的角度入射，同时振幅相等，偏振态相同，所形成干涉条纹的反衬度最高，此时成像系统的有效点扩展函数最优化；入射介质1的折射率越大，隐失干涉场的空间周期越短(空间频率越高)，其对应的调制点扩展函数中心瓣的半峰全宽越小，可能分辨更小的物体，但同时旁瓣的强度也增强，最终成像的分辨力受两者的共同制约。     

Computer Simulation of a 3-D Sensing System with Structured Illumination

A computer simulation system of three-dimensional sensing with structured illumination is presented. It includes the generation of deformed fringe patterns from 3-D shapes and the reconstruction of 3-D shapes, conversely. Some experimental results of the deformed fringe pattern and the reconstructed object shapes are presented. We have also discussed the effect of some major system parameters on the measurement results and considered how to correct these parameters according to the measurement result of the standard plane. Using this simulation system, the major system parameters: environmental conditions, measurement accuracy and algorithm evaluation of the 3-D shape measurement system based on PMP, FTP, SPM, etc., can be researched. © 1997 Elsevier Science Ltd. All rights reserved.     

The Problem of Subrayleigh Resolution in Interference Microscopy

A method allowing subwavelength resolution within the framework of optical interferometric microscopy is proposed. It is shown that overcoming the diffraction limit is not a necessary requirement. Generally speaking, subrayleigh resolution and overcoming the diffraction limit are basically different concepts. The method developed uses the all-parameter modulation of the light source and separation of the phase shifts generated due to different reasons. In this method, a topological phase technique is used to separate the phases and therefore determine the corresponding characteristics of an object. The approach developed provides for a way to create a new type of optical devices. Within the framework of a uniform measuring procedure these devices make possible determining both geometrical and material parameters of the object under study.     

荧光共焦并行探测显微成像系统设计及实验

将数字微镜器件加入到荧光显微成像系统中,代替传统共焦显微系统中的照明针孔,利用其调制特性,通过在数字微镜器件上加载不同图片,实现对光束的分割.对马铃薯细胞分别进行四通道、六通道、九通道的荧光细胞探测,同时采用平面反射镜作为样品,测试得到系统的深度响应曲线,分析了系统的分辨率.实验结果表明,数字微镜器件的加入实现了从点对点共焦成像变为多点并行共焦显微成像,提高了显微成像的探测速度,同时具有较高的分辨率.     

Detection of Single Quantum Dots in Model Systems with Sheet Illumination Microscopy

Single molecule detection and tracking provides at times the only possible method to observe the interactions of low numbers of biomolecules, inlcuding DNA, receptors and signal mediating proteins in living systems. However, most existing imaging methods do not enable both high sensitivity and non-invasive imaging of large specimens. In this study we report a new setup for selective plane illumination microscopy (SPIM), which enables fast imaging and single molecule tracking with the resolution of confocal microscopy and the optical penetration beyond 300 μm. We detect and report our instrumental figures of merit, control values of fluorescence properties of single nano crystals in comparison to both standard widefield configurations, and also values of nanocrystals in multicellular “fruiting bodies” of Dictyostelium, an excellent control as a model developmental system. In the Dictyostelium , we also report some of our first tracking of single nanocrystals with SPIM. The new SPIM setup represents a new technique, which enables fast single molecule imaging and tracking in living systems.     

10 nm Spatial Resolution Fluorescence Imaging of Single Molecules by Near-Field Scanning Optical Microscopy Using a Tiny Aperture Probe

We demonstrate high-resolution fluorescence imaging of single molecules using near-field scanning optical microscopy (NSOM) with a tiny aperture probe for two different wavelengths in visible range in the illumination mode of operation. The spatial resolutions obtained at both excitation wavelengths were almost the same and the highest resolution realized was about 10 nm. To discuss the achievable resolution in aperture NSOM, we also employed a computer simulation by the finite-difference time-domain method for various aperture sizes and wavelengths. The resolution of 10 nm is predicted to be contributed by the single peak of localized near-field light around the rim of the aperture.     

荧光谱线分布对共焦荧光显微镜分辨率的影响

研究了荧光谱线分布对共焦荧光显微镜中分辨率的影响,导出了荧光谱线分布的荧光功率传输函数、三维脉冲响应函数.数值计算了荧光谱线均匀分布的情况,结果表明：与采用单一中心荧光波长的分辨率比较,共焦荧光显微镜的横向分辨率、纵向分辨率随着荧光谱线范围和色散的增大而下降;谱线响应范围小的探测器有利于分辨率.     

High Resolution B-Type Delayed Fluorescence of Dibenzanthracene in PMMA

B-Type delayed fluorescence of 1,2,3,4-dibenzanthracene in PMMA was experimentally observed for the first time. Dibenzanthracene molecules were exerted in a two-step process. In the first step, an excited singlet S₁ is created, which undergoes intersystem crossing to T₁ then T-T absorption creates an excited triplet dibenzanthracene molecule, which returns to the first exerted singlet level by intersystem crossing. The recreated first excited singlet of dibenzanthracene decays back to the ground state by emitting this new type of delayed fluorescence.     

饱和激发下叶绿素荧光非线性变化及影响分析

叶绿素浓度是海洋初级生产力的重要指标之一,激光诱导荧光技术可以实现海水叶绿素浓度的快速测量。测量叶绿素浓度的传统激光诱导荧光原理,是利用叶绿素荧光与水体Raman散射的强度比值（I_F/R）进行反演,即叶绿素浓度n_chl＝CI_F/R,其中C为系统常量。这是依据叶绿素荧光685 nm、水体Raman散射强度都与激发光强呈线性关系。然而,该理论并没有考虑诱导荧光饱和现象的存在。当诱导激光强度达到一定程度后,685 nm荧光强度随激发光强非线性变化。另外,值得注意的是,水体Raman散射并不存在信号饱和现象。为了探讨饱和激发造成荧光非线性变化的影响,在激光诱导荧光技术测量叶绿素浓度的实验中,设计两种测量方案,即：不同激光功率诱导单一浓度样本的荧光测量,和固定激光功率时不同浓度样本的荧光测量。实验中利用Nd∶YAG三倍频激光355 nm激发获得叶绿素溶液的404 nm处 Raman散射和685 nm荧光。实验结果分为2部分进行讨论：（1）为了分析饱和激发造成荧光变化的非线性特性,通过调节激发光功率测量溶液的受激发射光谱,发现水体Raman散射强度与激发光强呈线性关系,而685 nm荧光强度出现饱和激发下的非线性变化。而且,随叶绿素浓度的增加,685 nm荧光的非线性趋势更为明显,Raman散射强度与激发光强的线性关系中斜率变小。数据分析表明,685 nm荧光数据拟合的4阶多项式和Raman散射效率值,可以定性地表征685 nm荧光的饱和程度。（2）考虑实际海洋激光雷达探测叶绿素浓度应用中存在饱和激发荧光非线性现象,为了分析荧光非线性对传统叶绿素浓度反演理论适用性的影响,在固定激发光强情况下对不同浓度叶绿素溶液的发射光谱进行测量。将激发光功率调节至52.00,80.70,132.10和197.30 mW·cm-2,获取相应激发光强下685 nm荧光与水体Raman散射的强度比值和叶绿素浓度之间的关系。实验表明,激发光强不变的情况下,685 nm荧光与水体Raman散射的强度比值,与叶绿素浓度仍满足线性关系。但是,在较高光强激发时,饱和激发造成的叶绿素荧光非线性变化,导致利用传统激光诱导荧光理论反演的叶绿素浓度值偏小。因此,需要对饱和激发下荧光非线性的影响进行修正,其关系为I_F/R=n_chl/C+C_F,修正值C_F不可忽略。另外,值得一提的是,修正关系中系统常量C随激发光强增加而增大。研究表明,饱和激发造成的荧光非线性,会对激光诱导荧光技术测量叶绿素浓度产生影响,但由于造成荧光非线性因素的复杂性,仅通过荧光数据拟合获得的多项式,无法定量说明其影响权重。然而,当激发光强不变时,可以实验测量获得基于激光诱导荧光原理的修正关系,从而准确反演叶绿素浓度。     

温度对大功率LED照明系统光电参数的影响

利用板上芯片封装chip-on-board(COB)技术封装大功率LED,比较分析其在不同散热器上的温度变化规律。研究了不同的热平衡温度对大功率LED光通量、电学参数的影响。在实验过程中,光通量、驱动电压、功率和发光效率都呈现出下降的趋势,并且最终稳定在其热平衡值。研究还发现:对于大功率LED照明系统,光通量、驱动电压、发光效率与散热器温度具有线性关系。在电源接通时,随着散热器温度的升高,LED的反向饱和电流迅速升高。通过线性拟合,得到大功率LED照明系统的光通量温度系数、驱动电压温度系数和光效温度系数。     

应用小波边缘检测技术分析荧光寿命显微像

研究了小波图象边缘检测技术在荧光寿命显微像分析中的应用，利用小波分析的多尺度特性进行了荧光寿命显微像的边缘提取，结果表明，这种方法非常有效。     

Spatial Resolution of Acoustic Microscopy in the Visualization of Interfaces inside a Solid

Acoustical Physics - The volumetric visualization of materials and inner boundaries of a joint is one of the fields in which long-focus high-frequency beams are used. Visualization is carried out...     

数字全息显微系统结构参量对再现像质的影响

为了获取高质量的数字全息显微再现像,分析了同轴相移数字全息显微系统结构参量对再现像质的影响.首先经过计算得出理想成像时,像空间物光波频率完全由物体面形结构频率和系统放大倍率决定.然后基于显微成像时物光波的所有频率分量都应该被有效记录的分析,得出在同轴相移系统中放大倍率必须使物光波的空间频率缩小到满足采样要求|并得出在相同的放大倍率下记录距离会影响有效记录的物体尺寸,记录器件离开像平面的距离越小,有效记录的物体尺寸越大,在像平面上时有效记录的物体尺寸最大.计算机仿真和光学实验都证明了上述结论的正确性.     

数字全息显微系统结构参量对再现像质的影响

为了获取高质量的数字全息显微再现像,分析了同轴相移数字全息显微系统结构参量对再现像质的影响.首先经过计算得出理想成像时,像空间物光波频率完全由物体面形结构频率和系统放大倍率决定.然后基于显微成像时物光波的所有频率分量都应该被有效记录的分析,得出在同轴相移系统中放大倍率必须使物光波的空间频率缩小到满足采样要求;并得出在相...     

复眼透镜阵列应用于均匀照明系统的特性研究

研究了复眼透镜阵列实现均匀照明的原理,模拟分析了复眼透镜的列数、入射光发散角、光源尺寸对照明效果的影响。仿真表明,入射光的发散角越小,光源尺寸越小,照明效果越好,而且适当地选取复眼透镜的列数能提高照明效果。     

Direct Observation of Lipid Domains in Free-Standing Bilayers Using Two-Photon Excitation Fluorescence Microscopy

The direct observation of temperature-dependent lipid phase equilibria, using two-photon excitation fluorescence microscopy on giant unilamellar vesicles (GUVs) composed of different lipid mixtures, provides novel information about the physical characteristics of lipid domain coexistence. Physical characteristics such as the shape, size, and time evolution of different lipid domains are not directly accessible from the traditional experimental approaches that employ either small and large unilamellar vesicles or multilamellar vesicles. In this review article, we address the most relevant findings reported from our laboratory regarding the direct observation of lipid domain coexistence at the level of single vesicles in artificial and natural lipid mixtures. In addition, key points concerning our experimental approach will be discussed. The unique advantages of the fluorescent probe 6-dodecanoyl-2-dimethylaminonaphthalene (LAURDAN) under two-photon excitation fluorescence microscopy is particularly addressed, especially, the possibility of obtaining information on the phase state of different lipid domains directly from the fluorescent images.     

应用遗传算法拟合偏振X射线荧光重叠谱

X射线荧光光谱分析由于受能量探测器分辨率的限制,谱线重叠干扰严重。不采用合适的重叠谱峰分解或曲线拟合技术,很难进行成分定性和定量分析。样品中的元素及谱线未知、背景基线不确定和模型初始参数不准确是曲线拟合中的最大困难。有多种算法可应用于光谱分析中的曲线拟合。文章将遗传算法应用于永磁材料偏振X射线荧光中的重叠谱分解,研究了进化策略对谱峰分解质量的影响,比较了遗传算法与传统算法的拟合结果。研究表明遗传算法在谱线严重重叠情况下仍具有较强的谱峰分解能力;群体初始化和进化策略的正确选择是该算法成功应用的关键;遗传算法具有全局搜索能力,对重叠谱峰的分辨能力优于标准Marquardt-Levenberg算法。     

Locating of Nucleic Acid Intercalators in Yeast Cells by Image Analysis Combined Fluorescence Microscopy

Intracellular distribution in the intact (not fixed) Saccharomyces cerevisiae cells of the nucleic acid intercalators (NAI) was studied using fluorescence microscopy combined with computer image analysis (ImageJ software, NIH, USA). Three NAI—the anthracycline anticancer drug doxorubicin (DR) along with the nucleic acid dyes ethidium (E) and 4′,6-diamidino-2-phenylindole (DAPI)—were used. Staining pattern and ImageJ quantitative analysis data provided evidence that all three NAI were located in the nuclei and in the mitochondria. DR and E, in contrast to DAPI, may be bound to not only DNA, but to the mitochondrial membranes as well. Experiments on the combined application of DR+DAPI and E+DAPI have shown potential competition of DAPI with DR and E for binding sites in the nuclear and mitochondrial DNA. With the approach presented herein, the yeast cells of S. cerevisiae can be used as a model for locating intracellular sites of the fluorescing nucleic acid intercalators. This model may be of help in designing new DNA-targeted drugs and in preliminary studies of their interaction with eukaryotic cells.     

基于正交调制的激光测距系统和解模糊方法

提出一种基于正交调制的激光测距系统,该方案与传统的二次混频方法相比,提高了相位测量精度并简化了系统的硬件设计,针对距离测量过程中的距离模糊问题,提出了一种基于超定方程组的距离解模糊算法,避免了对最优解的搜索,最后使用2台K60激光测距仪和3台基于本方案的测距仪在精度为0.18 mm的国家标准基线上进行对比试验,结果显示,测距时间少于1.8 s,平均测量误差在2 mm以内,在60 m的量程内测距标准差在1 mm以内,验证了该方案相比较传统的方法具有更高的测量精度和速度。