Simultaneous confocal recording of multiple fluorescent labels with improved channel separation

Confocal microscopes are often used to study specimens labelled with fluorophores. A commonly used method for simultaneous recording of the distribution of multiple fluorophores is to divide the fluorescent light emitted by the specimen into different wavelength regions using dichroic and bandpass filters. These different wavelength regions are then distributed to multiple detectors. However, the broad and overlapping spectra of commonly used fluorophores often result in considerable crosstalk between channels. A new technique, intensity-modulated multiple-beam scanning (IMS) microfluorometry, can be used to reduce this cross-talk substantially. The IMS technique is implemented with two laser beams of different wavelengths, intensity-modulated at different frequencies, which illuminate the specimen simultaneously. The two laser wavelengths predominantly excite one fluorophore each. Fluorescent light from the specimen is divided into two wavelength regions (red and green) which are detected by two photomultiplier tubes. The output signals from the photomultiplier tubes are connected to lock-in amplifiers. The effect of using modulated laser beams, in combination with lock-in amplifiers, is strongly to reduce cross-talk between the channels. The performance of the IMS technique using various types of specimen is compared with the results obtained using the conventional multi-detector method.     

An efficient spectroscopic detection system for cathodoluminescence mode scanning electron microscopy (SEM)

A detection system for analytical cathodoluminescence (CL) mode scanning electron microscopy (SEM) is described. This incorporates a cold stage, an efficient light collector, a monochromator and a photomultiplier and used the photon counting technique. The efficiency of the component subsystems was optimized and calibrated, and the performance of alternative light collecting and monochromating equipment is compared. The operation of the photon counter is discussed. The digital output of the photon counter was fed into a multichannel scaler and thence to a computer. This was used to correct the observed count rate with the calibrated spectral variations in the performance of the detection system. Spectra obtained at both room temperature and liquid nitrogen temperacure and monochromatic and panchromatic SEM micrographs are given as examples illustrating the value of this technique. The factors governing the performance of the system are discussed. The forms of noise in the signal and in the detection system are described and the means for minimizing, avoiding or correcting for them are dealt with. Sources of spurious signals in the SEM are treated.     

数字锁相技术在FEM仪器采集系统中的应用

针对频率域电磁法仪器采集系统实时处理数据的要求，在设计一个基于DSK6713的高精度24位A／D采集系统的硬件基础上研究数字锁相技术处理数据的应用。利用数字锁相技术获取一次场、二次场多频信号的同相分量和垂相分量，再利用专业软件将归一化PPM值转换成电导率和磁导率，继而成像获取地下信息。     

Improvement of the Metrological Performance Stability of a Dual-Beam Optical System

A dual-beam optical measuring system with a single photodetector is described. The operation principle of the system is based on modulation and lock-in detection of the working and reference radiation beams at multiple frequencies. A single photon-coupled pair is used in the system for generating synchronous signals. It is shown that the correct adjustment of the phase shifts of the clock signals of the lock-in amplifiers can considerably reduce the effect of the instability of the photon-coupled pair signal on the result of measurements.     

Ultrasonic characterization on sequences of CFRP composites based on modeling and motorized system

Composites are a material class for which nondestructive material property characterization is as important as flaw detection. Laminates of fiber reinforced composites often possess strong in-plane elastic anisotropy attributable to the specific fiber orientation and layup sequence when waves are propagating in the thickness direction of composite laminates. So the layup orientation greatly influences its properties in a composite laminate. It could result in the part being rejected and discarded if the layup orientation of a ply is misaligned. A nondestructive technique would be very beneficial, which could be used to test the part after curing and requires less time than the optical test. Therefore a ply-by-ply vector decomposition model has been developed, simplified, and implemented for composite laminates fabricated from unidirectional plies. This model decomposes the transmission of a linearly polarized ultrasound wave into orthogonal components through each ply of a laminate. Also in order to develop these methods into practical inspection tools, motorized system have been developed for different measurement modalities for acquiring ultrasonic signals as a function of in-plane angle. It is found that high probability shows between the model and tests developed in characterizing cured layups of the laminates.     

Multiscale iterative voting for differential analysis of stress response for 2D and 3D cell culture models

Focused ion beam-scanning electron microscope (FIB-SEM) tomography is a powerful application in obtaining three-dimensional (3D) information. The FIB creates a cross section and subsequently removes thin slices. The SEM takes images using secondary or backscattered electrons, or maps every slice using X-rays and/or electron backscatter diffraction patterns. The objective of this study is to assess the possibilities of combining FIB-SEM tomography with cathodoluminescence (CL) imaging. The intensity of CL emission is related to variations in defect or impurity concentrations. A potential problem with FIB-SEM CL tomography is that ion milling may change the defect state of the material and the CL emission. In addition the conventional tilted sample geometry used in FIB-SEM tomography is not compatible with conventional CL detectors. Here we examine the influence of the FIB on CL emission in natural diamond and the feasibility of FIB-SEM CL tomography. A systematic investigation establishes that the ion beam influences CL emission of diamond, with a dependency on both the ion beam and electron beam acceleration voltage. CL emission in natural diamond is enhanced particularly at low ion beam and electron beam voltages. This enhancement of the CL emission can be partly explained by an increase in surface defects induced by ion milling. CL emission enhancement could be used to improve the CL image quality. To conduct FIB-SEM CL tomography, a recently developed novel specimen geometry is adopted to enable sequential ion milling and CL imaging on an untilted sample. We show that CL imaging can be manually combined with FIB-SEM tomography with a modified protocol for 3D microstructure reconstruction. In principle, automated FIB-SEM CL tomography should be feasible, provided that dedicated CL detectors are developed that allow subsequent milling and CL imaging without manual intervention, as the current CL detector needs to be manually retracted before a slice can be milled. Due to the required high electron beam acceleration voltage for CL emission, the resolution for FIB-SEM CL tomography is currently limited to several hundreds of nm in XY and up to 650 nm in Z for diamonds. Opaque materials are likely to have an improved Z resolution, as CL emission generated deeper in the material is not able to escape from it.     

A computerized signal acquisition system for the quantitative evaluation of EBIC and CL data in a SEM

Electron beam-induced current (EBIC) and cathodoluminescence (CL) are widely used methods to obtain information about recombination properties of semiconducting materials and their defects on a micrometer length scale. In this article a computerized SEM (scanning electron microscope) setup is described, which enables us to perform simultaneous measurements of several signals and automatic temperature-dependent measurements. As examples for the performance of this system we present results obtained by simultaneous EBIC/CL experiments, allowing a reconstruction of the defect geometry. In a second example, the temperature dependence of the EBIC contrast is analyzed, introducing the method of EBIC spectroscopy.     

Precision metrology of integrated circuit critical dimensions using in situ differential scanning electron microscopy

A technique for performing precision measurements of critical dimensions (CD) on integrated circuit (IC) features using the scanning electron microscope (SEM) is presented. The method is based on obtaining true in situ differential SEM video signals of IC features. The technique is characterized by its high sensitivity to feature edges, and relative slopes. Results are presented on the application of this technique to SEM measurements of IC patterns in silicon, photoresist, and aluminum. The method affords the capability of accurate pattern alignment during SEM imaging preceding linewidth measurements.     

Simple technique for obtaining photoacoustic spectra corrected for the spectral variation of the source in single scan

A simple technique is presented to obtain normalized photoacoustic (PA) spectra corrected for the spectral variation of the source in a single scan. The input light beam is passed through the center of a dual slot chopper, which splits it into two chopped output beams at two different frequencies at a fixed ratio. The beams fall on the sample and the reference kept side-by-side in the same sample chamber. The PA signals are detected by a microphone and processed by two lock-in amplifiers tuned at two different frequencies. The technique is tested by recording the PA spectra of standard samples.     

Investigation of fluorescent lamp phosphors using the combined CL and EDS modes of the SEM

The phenomenon of cathodoluminescence (CL) potentially offers the ideal tool for studying the phosphor materials used in fluorescent lamps, since it can be used directly on processed or unprocessed powders, on coatings in tubes, or on sections cut from tubes. Using examples of both single component materials and multi-component blends, it is demonstrated how a relatively unsophisticated dispersive CL system attached to a scanning electron microscope (SEM) can be used expediently in the extensive study of such phosphors. These studies can be significantly enhanced when other complementary modes of the SEM (e. g. the energy-dispersive x-ray analysis facility) are combined with the CL mode. The strength of the combined technique lies in the major role it can play in materials and processing aspects of the powders themselves, in the processing of the lamps (e.g. by optimising such parameters as coating thickness, packing density etc.), and in diagnostic studies of poor materials or lamps (e.g. by locating rogue particles/components and identifying their origin). The technique also provides a convenient method of studying the temperature stability of selected phosphors.     

双路同瞬值12位数据采集器

适用于APPLEⅡ微型机的多路数据采集器通常是由模拟开关对多个模拟量分时切换输入的。本文介绍一个与APPLEⅡ微型机适配的双路同瞬时采集的12位数据采集器。该采集器尤为适用于双通路锁相放大器的正交矢量运算或双光路测量信号的处理。该采集器设计简单,成本低,性能可靠。本文介绍了这种数据采集器的硬件构成,与计算机APPLEⅡ适配的系统控制信号联接方法,给出了采集两路同瞬时值12位数据采集器软件编程的流程图。     

基于SR7265硬件的虚拟锁相放大器、频谱仪、阻抗计和半导体参数分析仪

锁相放大器是用来测量微弱信号的专用仪器.即使噪音上千倍于被测信号,通过锁相放大器,也能得到精确结果.随着数字信号处理技术在仪器中的应用,可编程仪器变得越来越灵活.结合虚拟仪器的技术,论文通过对SR7265的硬件重新编程,在同一个硬件上实现了虚拟锁相放大器、虚拟频谱仪、虚拟阻抗计和虚拟半导体参数分析仪等等功能.本虚拟仪器采用了4层的逻辑结构.同样的功能也在一个FPGA板上成功地部署.     

Application of a new detector for cathodoluminescence measurements in the wavelength range to 1.8 μm

The energetic gap structure of semiconductors or insulators can strongly be influenced by the local appearance of inhomogenities, impurities, dopants or vacancies. A high spatial resolution cathodoluminescence (CL) measuring technique makes it possible to investigate this gap structure via spectral analysis of the emitted CL. This can lead to a detailed knowledge of the local defect distribution. The wavelength range which could be detected by CL measurements has, up to the present, been limited to values less than 1 μm, because no detectors were available for higher wavelengths. By use of a new germanium detector, the measuring range could be extended to 1.8 μm. This makes it possible to analyse the CL properties, both of materials with small gap energies and of deep impurities. The detector properties which are important for CL measurements are presented. The efficiency of the detector is demonstrated by CL investigations of barium titanate ceramics and silicon. The results are discussed and compared to results obtained using conventional detectors.     

基于电感式传感器的金属颗粒材质识别及粒径估计

传统的电感式颗粒传感器输出的是电感或电压幅值的脉冲信号,本质为标量信号。可通过脉冲信号的正负性区分金属颗粒是磁性或非磁性,且只能在已知颗粒材质的情况下估计颗粒的粒径。但在含有多种金属颗粒的油液中,基于标量信号的颗粒识别方法将失效。为此,本文采用了一种双锁相放大电路,将颗粒产生的复数域信号转化为一对直流信号。提出一种基于模糊隶属度函数的信号处理方法,实现了在噪音干扰下多种颗粒的材质识别和粒径估计。本文搭建了三线圈传感器实验系统。利用五种金属颗粒构建了隶属度函数,并进行系统标定。最后选取两种颗粒对标定后的系统进行了验证。结果表明系统对颗粒材质的识别准确,粒径估计误差小于2%。     

基于MATLAB仿真的锁定放大器的研究

研究了锁定放大器的实现原理,并根据该原理用MATLAB的SIMULINK工具箱对锁定放大器进行了建模和仿真。仿真结果表明锁定放大器具有卓越的微弱信号检测能力。在实际测试中。所设计的锁定放大器也很好的完成了信号的检测和拾取。     

远距离处固体面内位移测量的研究

为了利用激光多普勒效应实现远距离处固体面内位移测量，研究了激光多普勒差动方法和锁相放大、同态滤波等技术，设计出能获得大的多普勒信号强度的差动光路，高灵敏度、大Q值的锁相放大器和能重构丢失信号的同态滤波器等。实验结果证明将由它们所组成的系统用于50m处固体的面内位移测量，效果令人满意，其相对误差约1％。     

An automated image alignment system for the scanning electron microscope

We have developed an automated image alignment system for the scanning electron microscope (SEM). This system enables specific locations on a sample to be located and automatically aligned with submicron accuracy. The system comprises a sample stage motorization and control unit together with dedicated imaging electronics and image processing software. The standard SEM sample stage is motorized in the X and Y axes with stepping motors which are fitted with rotary optical encoders. The imaging electronics are interfaced to beam deflection electronics of the SEM and provide the image data for the image processing software. The system initially moves the motorized sample stage to the area of interest and acquires an image. This image is compared with a reference image to determine the required adjustments to the stage position or beam deflection. This procedure is repeated until the area imaged by the SEM matches the reference image. A hierarchical image correlation technique is used to achieve submicron alignment accuracy in a few seconds. The ability to control the SEM beam deflection enables the images to be aligned with an accuracy far exceeding the positioning ability of the SEM stage. The alignment system may be used on a variety of samples without the need for registration or alignment marks since the features in the SEM image are used for alignment. This system has been used for the automatic inspection of devices on semiconductor wafers, and has also enabled the SEM to be used for direct write self-aligned electron beam lithography.     

Contrast effects using a two-detector system in low-voltage scanning electron microscopy

A system of two opposite Everhart–Thornley detectors A and B has formerly been applied in conventional SEM for electron energies between 5 and 20 keV to separate material, topographic and other types of contrast by sum and difference signals. This technique can also be used successfully for low-voltage scanning electron microscopy. The decreasing information depth with decreasing electron energy shows differences in the surface composition and contamination which cannot be observed beyond 5 keV. Also below 5 keV material and topographic contrast can be separated and increased by the A + B and A − B signals, respectively.     

Monte Carlo simulation of CL and EBIC contrasts for isolated dislocations

Electron beam-induced current (EBIC) and cathodoluminescence (CL) are widely used to investigate semiconductor materials and devices, particularly to obtain information on the recombination properties and the geometry of defects. This report describes a simple formulation of CL and EBIC contrasts based on the Born approximation of excess carrier density in the presence of a pointlike defect. Quantitative interpretation of the CL and EBIC images is often difficult because of a lack of accurate theory treating simultaneously both the details of the electron beam penetration in the semiconductor and the generation of EBIC and CL signals. To overcome this difficulty, the Monte Carlo approach to the phenomenon of the electron beam penetration in solids has been developed to calculate the CL and EBIC signals during a simulation of the electron trajectory. Results for an inclined dislocation in GaAs are presented.     

Simple synchronic detection at audio frequencies through a PC sound card

We present a simple personal computer based, synchronic detection system that emulates a lock-in amplifier at audio frequencies. The inputs of the sound card are used to acquire two sets of samples: the one to be measured, previously preamplified, and the reference. From the last one, two "quasiorthogonal" signals are derived to perform dual-phase detection. The procedure is fast and compares favorably with a benchtop lock-in amplifier. In the band from 100 Hz to 20 kHz we obtained average amplitude and phase errors below 1% and 0.1 degrees, respectively.