Three-dimensional automated nanoparticle tracking using Mie scattering in an optical microscope

The forward scattering of light in a conventional inverted optical microscope by nanoparticles ranging in diameter from 10 to 50nm has been used to automatically and quantitatively identify and track their location in three-dimensions with a temporal resolution of 200ms. The standard deviation of the location of nominally stationary 50-nm-diameter nanoparticles was found to be about 50nm along the light path and about 5nm in the plane perpendicular to the light path. The method is based on oscillating the microscope objective along the light path using a piezo actuator and acquiring images with the condenser aperture closed to a minimum to enhance the effects of diffraction. Data processing in the time and spatial domains allowed the location of particles to be obtained automatically so that the technique has potential applications both in the processing of nanoparticles and in their use in a variety of fields including nanobiotechnology, pharmaceuticals and food processing where a simple optical microscope maybe preferred for a variety of reasons.     

动态线偏振光散射纳米颗粒粒度测量法的研究与分析

多重散射是传统动态光散射法测量纳米颗粒溶液浓度上限受到限制的主要原因。为此文中提出了动态线偏振光散射纳米颗粒粒度测量法,通过改变颗粒入射光和散射光的偏振状态,降低颗粒间多重散射的影响。现利用Mie散射理论分析了入射光与散射光偏振状态之间的关系,并通过实验方法探知偏振光在散射介质中的传输特性,揭示了在动态光散射中使用垂直偏振光作为入射光的实验依据。最后对动态线偏振光散射颗粒测量法和传统光子相关光谱测量法进行了实验及分析,通过两种方法的比较,验证了上述理论的正确性。     

Microspectroscopic measurement of the optical properties of rat liver in the visible region

Microspectroscopy is used to investigate optical properties of haemoglobin-free perfused rat liver. Visible spectra of 20 μm diameter spot size were measured in transmission and/or reflection modes as a function of the thickness (< 1200 μm) of the liver-edge. Optical density (OD) in transmission mode increased with the increasing liver thickness, whereas in reflection mode OD decreased but became almost constant above a certain thickness (c. 600 μm) of the liver. The Kubelka-Munk (KM) two-flux model, with a minor modification, was applied successfully to the analysis of the changes in OD as a function of the thickness. This approach estimates the KM absorption coefficient (E_KM), KM scattering coefficient (S_KM) and effective penetration depth (δ_eff) of the liver. The optical properties were similar to reported values, obtained with different methods.     

Synthesis and optical properties research of gold nanoparticles with different morphologies

本文制备了不同形貌的金纳米颗粒,并对其形貌对光学性能的影响进行了研究。本文用还原法制备了不同粒径的金纳米颗粒,采用晶种生长法成功地制备出了星形、梭形和棒状的金纳米颗粒。颗粒的形貌和大小并采用投射电子显微镜(TEM)进行了表征,结果说明,本文成功制备出了不同形貌大小的金纳米颗粒。UV-Vis光谱和拉曼光谱仪对制备的颗粒的表征测试说明,不同形貌大小对颗粒有着不同的光学性能。拉曼光谱的结果说明,不同形貌大小的金纳米颗粒可以用作不同浓度分子的探针,对物质进行检测。     

Evanescent-wave microscopy: A simple optical configuration

Under suitable conditions, the region of the aqueous phase immediately adjacent to a glass-water interface can be selectively illuminated using the evanescent wave created when total internal reflection occurs at the interface. Objects in the aqueous phase away from the glass become effectively invisible, since the intensity of the evanescent wave decays exponentially with distance from the interface. Previous methods of generating evanescent waves for light microscopy have employed accessory light sources and optical components that directed the illumination on the specimen from one side. The asymmetric illumination creates a surprising orientation dependence of visibility for some specimens. Objects such as microtubules are totally invisible unless they are orientated nearly perpendicular to the direction of illumination. An explanation of this phenomenon is provided in terms of the geometry of diffraction of light by long thin objects. A simple method of achieving evanescent-wave illumination is described and shown to be useful in practice for biological specimens. In contrast to previously described methods, the present arrangement has the advantage of producing circularly symmetric illumination, and of utilizing only standard optical microscope components. The system has been used for imaging specimens both by light scattering and by fluorescence. It has proved useful for following the fragmentation of flagella into isolated microtubules, observing microtubules gliding over dynein adsorbed to a surface, and also for determining the arrangement of vinculin and alpha-actinin in cell-substratum attachment sites and termini of growing myofibrils in cardiac cells.     

A simple method allowing DIC imaging in conjunction with confocal microscopy

Current optical methods to collect Nomarski differential interference contrast (DIC) or phase images with a transmitted light detector (TLD) in conjunction with confocal laser scanning microscopy (CLSM) can be technically challenging and inefficient. We describe for the first time a simple method that combines the use of the commercial product QPm (Iatia, Melbourne Australia) with brightfield images collected with the TLD of a CLSM, generating DIC, phase, Zernike phase, dark-field or Hoffman modulation contrast images. The brightfield images may be collected at the same time as the confocal images. This method also allows the calculation of contrast-enhanced images from archival data. The technique described here allows for the creation of contrast-enhanced images such as DIC or phase, without compromising the intensity or quality of confocal images collected simultaneously. Provided the confocal microscope is equipped with a motorized z-drive and a TLD, no hardware or optical modifications are required. The contrast-enhanced images are calculated with software using the quantitative phase-amplitude microscopy technique ( Barone-Nugent et al., 2002 ). This technique, being far simpler during image collection, allows the microscopist to concentrate on their confocal imaging and experimental procedures. Unlike conventional DIC, this technique may be used to calculate DIC images when cells are imaged through plastic, and without the use of expensive strain-free objective lenses.     

Morphology, fluorescence properties, and their photothermal changes of poly(substituted thiophene) films revealed by near-field fluorescence microspectroscopy

The morphology and fluorescence spectrum of poly{3-[2-(N-dodecylcarbamoyloxy)ethyl]thiophene-2,5-diyl} film were examined with spatial resolution of 100 nm using near-field fluorescence microspectroscopy. Fluorescence spectra observed at protruding domains were blue-shifted compared with flat areas, and further blue-shift was observed there more appreciably by long-time irradiation via a near-field scanning optical microscope probe. It is considered that the polymer chains at the protruding domains take disordered conformations, in which conjugated lengths are shorter and further disordering can be induced more easily by irradiation compared with those in the flat areas.     

一种新型测量旋光色散的方法

提出了一种新的测量旋光色散的方法。该方法利用白光LED作为光源,经起偏棱镜、样品管和检偏棱镜后,复色偏振光经平面衍射光栅分光后,由线阵CCD进行采集,CCD的每个像元对应接收复色偏振光经分光后的某一波长的光信号,在检偏器旋转一周内,获得相应的光强数据,使用拟合算法,对每一个像元所采集的光强数据逐一进行处理,最终得出CCD像元所对应波长的旋光度,最后得到旋光色散数据。并利用标准石英管进行了实验研究,证明了这种测量方法的可行性。     

A method to correlate optical properties and structures of metallic nanoparticles

The optical response of individual nanoparticles is strongly influenced by their structures. In this report, we present a quick and simple pattern-matching based approach in which optical images of nanoparticles from localized surface plasmon resonance and single-molecule surface-enhanced Raman spectroscopy were used in conjunction with transmission electron microscopy for correlation of optical responses and the nanostructures of exactly the same nanoparticles or clusters of nanoparticles.     

Estimation of the section thickness and optical disector height with a simple calibration method

The distance between the upper and lower surfaces of a section (i.e. the section thickness) can be measured with a microcator or a shaft encoder. In the present report, an alternative simple method is described for estimating the section thickness where such equipment is not available. The basic principle of the method is based on a calibration method already described in the literature. The main difference is that it enables one to make more precise measurements. Provided that the calibration and measurements are made properly, this method can be used in estimating the section thickness, optical disector heights, and in particular in the determination of the thickness sampling fraction for the optical fractionator.     

用于镀制超薄银膜的一种有效的监控方法

银经常被应用于制备诱导增透滤光片,在这种宽带滤光片中,银膜的厚度一般在几个纳米,这样使得精确监控银的厚度很困难。提出一种简单易行的改进方法,通过在蒸发源上方加一个调速挡板,结合晶振膜厚仪,可以获得±0.4nm以内控制精度的超薄银膜。     

振幅调控双曲余弦高斯涡旋光束的聚焦特性

研究了双曲余弦高斯涡旋光束不同振幅参量对光场分布的影响。在相同光涡旋的情况下改变振幅的大小,在焦平面上场强分布变化明显。涡旋光束的拓扑数对光束的聚焦特性存在明显影响,不同的拓扑数会引起聚焦区域光强分布变化,另外也产生一些新的多焦点阵列和多重菱形聚焦图案。     

流动腐蚀方式对石英光纤湿腐蚀特性的影响

为了获得光滑的腐蚀光纤表面,本文从光纤腐蚀的传质及动力学特性出发,设计了一种流动腐蚀光纤装置.采用质量百分比浓度为12.5％的氢氟酸(HF)溶液,研究了石英光纤的组成成分、腐蚀剂温度和流速对腐蚀速率以及腐蚀后光纤表面形貌的影响.实验结果及理论分析表明:光纤腐蚀速率和表面粗糙度受化学反应速率和传质速率控制;由于光纤纤芯与包层成分不同,导致纤芯腐蚀速率高于包层腐蚀速率;在静态腐蚀条件下,腐蚀速率随温度呈非线性增长,且腐蚀后光纤表面粗糙;在流动腐蚀条件下,光纤腐蚀速率提高,并与温度呈线性关系,腐蚀后光纤表面粗糙度随流速的增加呈现出先减小后增大的趋势;在流速为0.75 L/min时,获得了光滑的腐蚀光纤表面.     

内表面污染物的低相干光干涉检测法

为检测透明容器内表面是否存在污染物,建立了基于容器壁内表面反射的低相干光干涉测量系统。实验采用宽带低相干半导体激光光源和改进的迈克尔逊干涉仪。将测量臂光路对准透明容器,并调节参考臂的光程,使容器内表面反射光的光程与参考臂反射光的光程相等,并出现干涉现象,捕捉并记录此动态干涉信号。经MATLAB软件处理并提取此信号,根据干涉光强度的不同即可辨别透明容器内是否存在污染物。该检测法可为生物医学、食品质检等领域检测透明容器内表面污染物提供一种方便快捷的新方法。     

微粗糙光学表面与多个镶嵌粒子差值散射场特性

本文基于时域有限差分算法,研究了微粗糙光学表面与多个镶嵌粒子的差值场光散射问题。将光学基片视为微粗糙光学表面,利用蒙特卡洛方法解决了光学表面存在粗糙度的问题,并将差值场散射理论加入到计算模型中,更好地分析了缺陷粒子的散射特性,将计算区域划分成上下两个半空间,建立了微粗糙光学表面与镶嵌多体粒子复合散射模型,并与矩量法计算结果比较验证了理论的有效性。运用此模型分析了入射角、镶嵌粒子尺寸、粒子间距、粒子个数等物性特征对微粗糙光学表面与镶嵌多体粒子差值散射场的影响。实验结果表明:在一定激光入射角下,以相同回波探测角度间距20°对光学表面进行测量能够有效地检测出缺陷粒子。本文结果为光学无损检测、光学薄膜、微纳米结构的光学性能设计等提供了理论依据和技术支持。     

扭叶罗茨轮的简易数显加工

扭叶罗茨轮，因加工困难，影响推广。经分析．提出方法只需将2坐标数显装置增加到普通铣床便可加工扭叶罗茨轮。     

圆锥量规锥度的高精度光学干涉法测量

针对超精密数控机床对圆锥量规锥度高精度测量的需求,研究了一种将“大数”和“小数”相结合,准确测量圆锥量规锥度的方法.介绍了锥度测量系统的基本原理及组成,给出了实验测量结果,分析了测量系统的不确定度.该测量方法利用最大分度间隔误差为0.10〞的多齿分度台构成高准确度分度系统保证测量“大数”部分的准确性,采用激光偏振干涉装...     

谱域光学相干层析成像量化技术及其在生物组织定量分析中的应用

研究了谱域光学相干层析(SDOCT)成像系统的量化技术,通过量化OCT图像来获得生物组织内部的信息特征对组织光学散射特性进行定量分析.给出并讨论了单次散射模型,具有轴向点散射函数(PSF)的单次散射模型和多次散射模型,利用平均A-scan算法和非线性最小二乘法曲线拟合,研究不同浓度IntralipidTM溶液的散射特性.实验显示,IntralipidTM溶液的散射系数与浓度在1％～10％间基本呈线性关系,验证了具有PSF的单次散射模型比较适用于本文的谱域光学相干层析成像系统.利用该模型对小鼠的新鲜肝脏进行量化研究,得到小鼠新鲜肝脏在波长λ.为1550 nm时的散射系数为8.9 mm-1.本文的研究为该项技术今后在临床医学上对各种疾病的诊断和治疗奠定了基础.     

光学表面冗余粒子在离轴及斜入射高斯波束作用下的辐射力

针对光学表面冗余粒子的清除问题,通过离轴高斯波束的球矢量波束展开方法,研究了介质球对离轴及斜入射高斯波束的散射特性;基于得到的结果结合连带勒让德函数的正交递推关系推导了离轴高斯波束对介质球的横向和轴向辐射力的解析表达式,并重点分析了离轴距离和高斯波束斜入射角度分别对轴向辐射力和横向辐射力的影响。结果表明,离轴入射时,在束腰负半轴2μm处出现一个总力极大值,过束腰零点后5μm处出现一个反向次大值点;高斯波束斜入射时,辐射力在束腰中心前后各出现一个极大值,随着入射角的增大,曲线整体下移减小,辐射力极大值所对应的Z0位置固定。得到的结论可用于无损检测工程中,即通过调节高斯波束束腰位置可提高对冗余粒子驱逐和控制的效率。