回束光导摄象管的特性与应用

回束光导摄象管(RBV)是近期发展起来的一种具有特殊性能的摄象管.4¹/2吋 RBV 管的突出特性包括高的孔径响应.高的信号增益,宽的动态范围和大容量的靶面.管子的电学设计参数与其他直读型和回束读出型的成象器件比较,指出了 RBV 管是一个本质上性能要高得多的器件.为了提供更高的灵敏度,正在发展一种带纤维光学耦合象增强器的回束管.RBV 管的运用包括图象传感和静电贮存.已证明总的分辨率可达100对线/mm(每图象高10000电视行).当100对线/mm时,RBV 管的性能可赶上或超过高分辨率胶片的性能.特别是在低对比度的情况下.对稳态光学曝光,读出可以连续进行,或者对于快门曝光或电学记录组成的断续输入,读出可以接近实时完成.对不连续输入,信息可以用慢速扫描单帧读出,或者为了在电视监示器上显示,信息可以用快速多帧读出.在多帧读出方式中,可以得到长达约一分钟的连续的高质量显示.读出传递函数(γ)可以通过电子学来控制.全部读出方式皆允许通过光栅控制和可变焦距进行电子放大.RBV 管好的性能和高的灵活性应能广泛地用在侦察系统,光学和电学贮存及扫描变换.数据取回和发送以及信号处理上.     

柔性制造系统切削稳定性在线监测技术的研究

主要讨论柔性制造系统(FMS)切削稳定性的在线监测方法,在研究FMS切削过程中振动信号变化规律的基础上^[1],提出了用μ_a=E(a)/F(a)作为FMS切削稳定性的监测参数,并进行了理论分析和试验验证.     

宽光谱长工作距弱荧光信号检测显微物镜设计

针对癌细胞突变基因诱导荧光信号弱、光谱覆盖范围宽、现有显微镜不能检测等局限,本文设计了光谱波段为450~800nm、数值孔径为0.95的荧光显微物镜。物镜采用++-结构,因宽光谱、大数值孔径像差校正难度大、透镜片数多、装调困难,前组设置成敏感组分,承载物镜装调的调校功能,承担90%以上光焦度;中间组为弱光焦度组分,用于校正大数值孔径下的二级光谱,显著降低了二级光谱校正元件的加工难度;后组为负光焦度组分,用于平像场和增大物镜的工作距离。物镜的设计参数为:总长58mm、工作距0.21mm、视场0.625mm、倍率40×、数值孔径0.95,结果表明:其像质接近衍射极限,畸变小于0.2%,满足多种癌细胞突变基因的弱信号生物监测设计要求。     

利用Monte Carlo模拟技术研究OCT图像对比度

建立了光在生物组织中传播的模型,利用MonteCarlo模拟技术快速计算和可移植性的优点,研究了OCT图像对比度与显微物镜数值孔径、焦距深度、时间门参数的关系,并给出它们的关系曲线.通过在所建立的模型基础上加入透镜透过率函数和光学传递函数,弥补了以往程序只能模拟光在生物组织中传播行为的缺点.该模型不仅可以分析生物组织图像与生物光学特性的关系,而且还可以指导OCT结构的完善和创新.模拟结果表明:在构建OCT时,参考臂与样品臂的这两个显微物镜的数值孔径越大,生物组织的采样深度越浅,处理信号的时间门宽度越小(但时间门宽度不能小于激光脉冲时间),混浊生物组织图像对比度越好.     

CCD图像存贮与显示系统

本文介绍了在合成孔径雷达光学处理器上所加的一套CCD图像存贮与显示系统.该系统采用线阵CCD器件,对运动的图像采样,其输出信号经A/D变换后,在行同步逆程期间将数字信号逐行写入存贮器中去;在行同步正程期间再将数据读出,经假彩色编码及D/A变换后即可在彩色监示器上显示出滚动的假彩色图像.该系统除了能够即时观察图像之外,还为进一步数字图像处理提供了方便.     

航空底片测试仪的恒张力数字控制系统研制

在航空底片测试仪的运动过程中,底片张力的变化对底片的位移和定位精度影响很大,从而导致测试结果的稳定性.在对测试仪传动机构的运动学和动力学分析的基础上,建立了全数字闭环控制模型.为了实现张力的实时采样和控制,采用数字信号处理技术(DSP)控制底片在运动过程中的张力,使张力趋于设定的最佳值.使得测试仪获取稳定的、真实的图像数据,为正确测评航空感光胶片成像质量提供了基础.实验结果表明底片平均移动速度最高可达到23m/min;张力设定范围是F_min=0N,F_max=30N;张力变化控制量为ΔF=±0.05N;重复定位精度为±10μm;满足了设计要求.     

铝合金搅拌摩擦焊液冷流道承压力学模型

为了实现液冷组件的可靠性设计,并且缩短液冷流道结构的设计周期,文中在通过力学仿真计算获得大量仿真数据的基础上,探索了流道承压共性规律,明确了液冷流道承压最大应力响应的影响因素及影响规律。结果表明,搅拌摩擦焊液冷流道内部的最大等效应力与盖板厚度呈反比,与盖板承压宽度的平方呈正比,与内压呈正比,在此基础上,建立了液冷流道承压力学模型 σ = KPL ² H ^-1,其中,对于铝合金液冷组件,K=0. 243 mm^-1。该承压力学模型为液冷组件提供了力学设计依据,根据液冷组件流道内压可以快速明确其几何尺寸,大大提高了设计效率,且广泛适用于采用其他焊接方法制造的同结构类型的液冷组件。     

新型光子纳米射流器件的设计

光子纳米射流是一种高强度,极窄的亚波长电磁场区域,它是由介电微球或微柱体的Mie散射对电磁场的聚焦作用产生的。光子纳米射流广泛应用于激光加工、纳米光刻、光学高密度存储以及超分辨率显微镜。从径向偏振光的角度出发,使用一种介电圆环结构对光束进行聚焦,由于径向偏振光在焦点区域可以产生较强的纵向场,通过优化圆环的尺寸、折射率以及与物镜焦点的相对位置,可以得到超过90%光束质量的纵向光子纳米射流,而且强度相比于未使用圆环时可以提高约一个数量级,并在高折射率下可以获得半高全宽小于衍射极限尺寸的光斑,因此该结构预计可以在粒子加速、光镊以及拉曼光谱学中有所应用。     

固溶及时效温度对TC18 组织性能的影响

固溶及时效处理是实现钛合金强化的关键工艺,文中采用不同的固溶及时效温度对TC18钛合金进行了强化处理,对处理后合金的显微组织和性能进行了对比分析。在固溶过程中,随着温度的升高,亚稳β相生成量增加,使得后续时效过程的次生α相析出量增加,从而提高了合金强度,但塑性降低;在时效过程中,随着温度的升高,初生α相晶粒长大,次生α相析出量减少,合金强化效果降低而塑性提高。通过控制固溶及时效温度,调整初生α相与次生α相之间的数量及尺寸关系,可以达到调整合金性能的目的。     

用宽带超表面产生阵列贝塞尔光束

通过在空间光调制器（SLM）上加载相位图或通过光刻加工微型圆锥状结构可以产生贝塞尔光束阵列。然而,典型空间光调制器具有比波长大一个数量级的像素尺寸,这限制了相位梯度的可用范围,用光刻法加工的微型锥透镜的顶端不是标准的圆锥,这影响了贝塞尔光束的质量。为了克服这些缺点,将复杂的相位图加载到电介质超表面上,设计了一种可以产生阵列贝塞尔光束（在波长700 nm处,NA=0.3）的超表面器件。该器件可以宽波段工作,其单元结构在波长580~800 nm范围内的偏振转换效率均超过57%。利用时域有限差分算法（FDTD）对该器件（厚度为380 nm,直径仅为40 μm）进行了仿真,所产生的阵列光束都垂直于超表面器件。所提出的阵列贝塞尔光束发生器具有纳米级别的厚度和几十微米的直径,这对于未来的集成光学领域具有很大的应用前景。     

A procedure to determine the correct thickness of an object with confocal microscopy in case of refractive index mismatch

A difference in refractive index (n) between immersion medium and specimen results in increasing loss of intensity and resolution with increasing focal depth and in an incorrect axial scaling in images of a confocal microscope. Axial thickness measurements of an object on such images are therefore not exact. The present paper describes a simple procedure to determine the correct axial thickness of an object with confocal fluorescence microscopy. We study this procedure for a specimen that has a higher refractive index than the immersion medium and with a thickness up to 100 µm. The measuring method was experimentally tested by comparing the thickness of polymer layers measured on axial images of a confocal microscope in case of a water–polymer mismatch to reference values obtained from an independent technique, i.e. scanning electron microscopy. The case when the specimen has a lower refractive index than the immersion medium is also shown by way of illustration. Measured thickness data of a water layer and an oil layer with the same actual thickness were obtained using an oil-immersion objective lens with confocal microscopy. Good agreement between theory and experiment was found in both cases, consolidating our method.     

Blind deconvolution of 3D fluorescence microscopy using depth‐variant asymmetric PSF

The 3D wide‐field fluorescence microscopy suffers from depth‐variant asymmetric blur. The depth‐variance and axial asymmetry are due to refractive index mismatch between the immersion and the specimen layer. The radial asymmetry is due to lens imperfections and local refractive index inhomogeneities in the specimen. To obtain the PSF that has these characteristics, there were PSF premeasurement trials. However, they are useless since imaging conditions such as camera position and refractive index of the specimen are changed between the premeasurement and actual imaging. In this article, we focus on removing unknown depth‐variant asymmetric blur in such an optical system under the assumption of refractive index homogeneities in the specimen. We propose finding few parameters in the mathematical PSF model from observed images in which the PSF model has a depth‐variant asymmetric shape. After generating an initial PSF from the analysis of intensities in the observed image, the parameters are estimated based on a maximum likelihood estimator. Using the estimated PSF, we implement an accelerated GEM algorithm for image deconvolution. Deconvolution result shows the superiority of our algorithm in terms of accuracy, which quantitatively evaluated by FWHM, relative contrast, standard deviation values of intensity peaks and FWHM. Microsc. Res. Tech. 79:480–494, 2016. © 2016 Wiley Periodicals, Inc.     

Utilizing confocal microscopy to measure refractive index of articular cartilage

This study proposes a method for measuring the refractive index of articular cartilage within a thin and small specimen slice. The cartilage specimen, with a thickness of about 50 μm, was put next to a thin film of immersion oil of similar thickness. Both the articular cartilage and immersion oil were scanned along the depth direction using a confocal microscope. The refractive index mismatch between the cartilage and the immersion oil induced a slight axial deformation in the confocal images of the cartilage specimen that was accurately measured by a subpixel edge‐detection‐based technique. A theoretical model was built to quantify the focal shift of confocal microscopy caused by the refractive index mismatch. With the quantitative deformations of cartilage images and the quantified function of focal shift, the refractive index of articular cartilage was accurately interpolated. At 561 nm, 0.1 MPa and 20 °C, the overall refractive index of the six cartilage plugs was 1.3975 ± 0.0156. The overall coefficient of variation of all cartilage specimens was 0.68%, which indicated the high repeatability of our method. The verification experiments using distilled water showed a minimal relative error of 0.02%.     

Refractive-index-induced aberrations in two-photon confocal fluorescence microscopy

The effect of refractive index mismatch on the image quality in two-photon confocal fluorescence microscopy is investigated by experiment and numerical calculations. The results show a strong decrease in the image brightness using high-aperture objectives when the image plane is moved deeper into the sample. When exciting at 740 nm and recording the fluorescence around 460 nm in a glycerol-mounted sample using a lens of a numerical aperture of 1·4 (oil immersion), a 25% decrease in the intensity is observed at a depth of 9 μm. In an aqueous sample, the same decrease is observed at a depth of 3 μm. By reducing the numerical aperture to 1·0, the intensity decrease can be avoided at the expense of the overall resolution and signal intensity. The experiments are compared with the predictions of a theory that takes into account the vectorial character of light and the refraction of the wavefronts according to Fermat's principle. Advice is given concerning how the effects can be taken into account in practice.     

Resolution in light microscopy studied by computer simulation

Microscopic resolution can be characterized by K = x NA/δ, where x is the distance between two objects, or the interval of a grating, just resolved with light of wavelength λ and an objective of aperture NA. Using a computer simulation of imaging the following K values were obtained on the Sparrow resolution criterion for line and grating objects and various imaging methods (figures for the Rayleigh criterion, which assumes a finite contrast-sensitivity of the light detector, are in parentheses). Several results appear to be novel. Due to limitations discussed in the text some data are only approximate. With a grating object K is 1.0 (1.0) for axial coherent illumination, 0.5 (0.5) for coherent illumination at an obliquity NA_obi which just enters the objective aperture, 0.5 (about 0.53) for incoherent illumination, 0.5 (about 0.52) for illumination with a condenser aperture NA_c equal to NA, 0.5 (about 0.515) for transmitted-light confocal scanning, and 0.25 (about 0.38) for fluorescent confocal scanning. If the object consists of two parallel lines K is about 0.68 (0.71) for axial coherent illumination, about 0.44 (0.5) for incoherent illumination, 0.375 (about 0.48) for optimal partially coherent illumination in which NA_c may exceed NA, 0.44 (0.48) for transmitted-light confocal scanning, and 0.32 (0.41) for fluorescent confocal scanning. For inter-object distances of 1, 1.5 and 2 wavelengths, respectively, NA_c values of about 0.69, 0.5 and 0.375 gave optimal contrast and resolution irrespective of NA. The practice of setting NA_c to about two-thirds of the NA of a high-power objective is supported by the fact that a condenser aperture of about 0.69 gives excellent or optimum resolution and contrast for most inter-object distances and objective apertures tested, although with some distances and apertures reducing NA_c improved contrast slightly. The rule (sometimes attributed to Abbe) that resolving power is proportional to the mean of NA and NA_c is correct for oblique coherent illumination in the case of a grating object, provided NA_obl does not exceed NA. In the case of two isolated objects the rule is only approximately correct, but applies even if NA_obl is greater than NA. Coherent light at an obliquity of 0.5λ/x introduces a half-wavelength phase difference between two objects and permits their resolution (with perhaps an incorrect apparent inter-object distance) even with objective apertures approaching zero. In confocal scanning the width of the scanning spots has only a moderate effect on resolution, and two objects can sometimes be resolved with scanning spots wider than the inter-object distance provided the lens apertures are neither too small nor too large.     

Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index

Accurate distance measurement in 3D confocal microscopy is important for quantitative analysis, volume visualization and image restoration. However, axial distances can be distorted by both the point spread function (PSF) and by a refractive‐index mismatch between the sample and immersion liquid, which are difficult to separate. Additionally, accurate calibration of the axial distances in confocal microscopy remains cumbersome, although several high‐end methods exist. In this paper we present two methods to calibrate axial distances in 3D confocal microscopy that are both accurate and easily implemented. With these methods, we measured axial scaling factors as a function of refractive‐index mismatch for high‐aperture confocal microscopy imaging. We found that our scaling factors are almost completely linearly dependent on refractive index and that they were in good agreement with theoretical predictions that take the full vectorial properties of light into account. There was however a strong deviation with the theoretical predictions using (high‐angle) geometrical optics, which predict much lower scaling factors. As an illustration, we measured the PSF of a correctly calibrated point‐scanning confocal microscope and showed that a nearly index‐matched, micron‐sized spherical object is still significantly elongated due to this PSF, which signifies that care has to be taken when determining axial calibration or axial scaling using such particles.     

基于后向光散射的颗粒测量技术研究

颗粒粒径和颗粒折射率是光散射颗粒测量技术中的重要参数。为了实现颗粒粒径的测量及其分档,在广义Mie理论基础上,分析了颗粒粒径及折射率对后向散射光能分布的影响,并得到了后向散射光能分布随颗粒粒径及折射率呈周期变化规律。实验验证结果表明,后向散射光能与颗粒粒径及颗粒浓度有关。研究结果可为后续的颗粒测量研究提供参考。     

3D deconvolution of spherically aberrated images using commercial software

Refractive index mismatch between the specimen and the objective immersion oil results in spherical aberration, which causes distortion and spreading of the point spread function, as well as incorrect readings of the axial coordinates. These effects have to be taken into account when performing three-dimensional restoration of wide-field fluorescence images. By using objects with well-defined geometry (fluorescently stained Escherichia coli or actin filaments) separated from a cover slip by a layer of oil with known refractive index, we investigated the accuracy of three-dimensional shape restoration by the commercial programs Huygens and Autoquant. Aberration correction available in the software dramatically reduced the axial blur compared to deconvolution that ignored the refractive index mismatch. At the same time, it failed to completely recover the cylindrical symmetry of bacteria or of actin fibres, which showed up to a three to five times larger width along the optical axis compared to the lateral plane. The quality of restoration was only moderately sensitive to the exact values of the specimen refractive index but in some cases improved significantly by assuming a reduced NA of the objective. Because image restoration depends on the knowledge of the vertical scale, we also performed detailed measurements of the axial scaling factor and concluded (in agreement with some previous authors) that scaling is adequately described by the simple paraxial formula, even when high-NA oil-immersion objectives are used.     

High‐resolution wide‐field microscopy with adaptive optics for spherical aberration correction and motionless focusing

Live imaging in cell biology requires three‐dimensional data acquisition with the best resolution and signal‐to‐noise ratio possible. Depth aberrations are a major source of image degradation in three‐dimensional microscopy, causing a significant loss of resolution and intensity deep into the sample. These aberrations occur because of the mismatch between the sample refractive index and the immersion medium index. We have built a wide‐field fluorescence microscope that incorporates a large‐throw deformable mirror to simultaneously focus and correct for depth aberration in three‐dimensional imaging. Imaging fluorescent beads in water and glycerol with an oil immersion lens we demonstrate a corrected point spread function and a 2‐fold improvement in signal intensity. We apply this new microscope to imaging biological samples, and show sharper images and improved deconvolution.     

Aberrations in confocal fluorescence microscopy induced by mismatches in refractive index

The effect of refractive-index mismatch, as encountered in the observation of biological specimens, on the image acquisition process in confocal fluorescence microscopy is investigated theoretically. The analysis takes the vectorial properties of light into account and is valid for high numerical apertures. Quantitative predictions on the decrease of resolution, intensity drop and shift of focus are given for practical situations. When observing with a numerical aperture of 1·3 (oil immersion) and an excitation wavelength of 514 nm the centre of the focus shifts 1·7 μm per 10 μm of axial displacement in an aqueous medium, thus yielding an image that is scaled by a factor of 1·2 in the axial direction. Furthermore, it can be expected that for a fluorescent plane 20 μm deep inside an aqueous medium the peak intensity is 40% less than for a plane which is 10 μm deep. In addition, the axial resolution is decreased by a factor of 1·4. The theory was experimentally verified for test samples with different refractive indices.