A real‐time image dynamic range compensation for scanning electron microscope imaging system

This paper presents the development and implementation of a real‐time dynamic range compensation system for scanning electron microscope (SEM) imaging applications. Compared with conventional automatic brightness contrast compensators that are based on the average image or pixel intensity level, the proposed system utilizes histogram‐profiling techniques to compensate continuously the dynamic range of the processed video signal. The algorithms are implemented in software with a frame grabber card forming the front‐end video capture element. The proposed technique yields better image compensation compared with conventional methods.     

Bridge vibration logarithmic decrement estimation at the presence of amplitude beat

Logarithmic decrement is one of the key dynamic parameters characterizing bridge status. It is usually estimated from vibration records applying digital signal processing. Among the processing procedures band-pass filtering is commonly used to extract spectral components of interest. We analyze the case when two very close spectral components are present in the acceleration records. We show that construction of the filter is non-trivial in such a situation and failing to choose optimal filter’s parameters leads to significant logarithmic decrement estimation errors. Therefore we propose a method that does not require band-pass filtering but utilizes knowledge of amplitude beat presence. Simulation of the method indicated errors by an order less than using filtering.     

面向微纳材料的激光扫描二维成像系统

在对微纳材料光学特性表征中,需要获得分辨率更高的波长和强度的荧光图像。普通的显微镜无法满足测试的要求,因此将普通的成像显微镜、光谱仪以及纳米移动台组成激光扫描显微镜成像系统,并利用LabVIEW开发了一套完整的集二维扫描采集与信号图像处理一体的系统上位机软件。扫描采集过程使用了低通滤波等数字信号处理方法消除光谱仪信号噪声的影响。利用本系统测量硒化镉纳米带、单层二硫化钼得到了荧光强度图像以及荧光峰值波长图像,能分辨出最小波长为0.03 nm的荧光。将采集长度与实际长度进行比较并分析荧光强度差异,取得了较好的效果。     

Spectrally resolved cathodoluminescence analyses in the optical near-field

By implementing a scanning near-field optical microscope into the specimen chamber of a scanning electron microscope, cathodoluminescence can be locally detected in the optical near-field. The achievable spatial resolution in this set-up is only limited by the size of the aperture in a coated fibre probe and its separation from the sample, rather than by the energy dissipation volume of the primary electrons and diffusion processes of excess carriers inside the specimen. We demonstrate how electronically active defects in polycrystalline diamond can be distinguished and localized with sub-wavelength lateral resolution by spectral filtering of the cathodoluminescence signal.     

Zero-loss energy filtering under low-dose conditions using a post-column energy filter

Electron cryomicroscopy combined with energy filtering can be performed under low-dose conditions using a post-column energy filter. The microscope combined with the filter is set up such that it can be used with similar ease as a conventional microscope, the main difference being that all filter and microscope control is performed through a central computer and images are recorded with a cooled slow-scan CCD camera. The microscope can also still be used for regular imaging on film as without the filter. Owing to the 18 times post-magnification of the filter, the microscope normally has to be operated at a small magnification, e.g. 3000×, and the beam has to be contracted to a small spot, e.g. 5 mm, in the plane of the microscope viewing screen. Computer control allows one to perform a variety of tasks automatically, such as autofocusing, thickness measurements, most-probable-loss imaging, CCD spot-scanning and tomography. The gain in contrast due to zero-loss energy filtering is analysed using visual inspection, power spectra and Fourier ring correlation. The thickness range for ice-embedded specimens in which a filter at 120 kV is most useful appears to be between 100 and 300 nm.     

Spectrally resolved cathodoluminescence analyses in the optical near-field

By implementing a scanning near-field optical microscope into the specimen chamber of a scanning electron microscope, cathodoluminescence can be locally detected in the optical near-field. The achievable spatial resolution in this set-up is only limited by the size of the aperture in a coated fibre probe and its separation from the sample, rather than by the energy dissipation volume of the primary electrons and diffusion processes of excess carriers inside the specimen. We demonstrate how electronically active defects in polycrystalline diamond can be distinguished and localized with sub-wavelength lateral resolution by spectral filtering of the cathodoluminescence signal.     

Application of the "imaging plate" to TEM image recording

The "imaging plate" is a highly sensitive image recording plate for X-ray radiography, which is coated with photo-stimulable phosphor. The imaging plate is exposed to electrons in a transmission electron microscope. Its fundamental properties (sensitivity, dynamic range and sharpness) have been estimated in detail. Also, the image quality of the imaging plate for some specimens in a transmission electron microscope has been estimated. As a result, it has been ascertained that the imaging plate has superior properties and high practicability as an image recording material in a transmission electron microscope.     

显微高光谱成像系统的设计

设计出一种基于棱镜 光栅 棱镜组合分光方式的显微高光谱成像实验系统.系统根据推帚式成像光谱仪的原理进行设计,采用棱镜 光栅 棱镜组合元件在后光学系统进行光谱分光,利用高精度载物台自动装置驱动样品进行推扫成像,选用PCI总线作为数据采集的微机接口.整个系统由显微镜、分光计、面阵CCD相机、载物台自动装置以及数据采集与控制模块等几部分组成.系统的光谱范围从400nm到800nm,120个波段,光谱分辨率优于5nm,空间分辨率大约1μm.该系统具有直视性、光谱分辨率高、结构紧凑、成本低等优点;不仅能够提供微小物体在可见光范围的单波段显微图像,而且能够获得图像中任一像素的光谱曲线,实现了光谱技术和显微成像技术的结合,成功的将成像光谱技术应用到显微领域,可广泛应用于临床医学、生物学、材料学、微电子学等学科领域.     

Digital-signal-processor-based dynamic imaging system for optical tomography

In this article, we introduce a dynamic optical tomography system that is, unlike currently available analog instrumentation, based on digital data acquisition and filtering techniques. At the core of this continuous wave instrument is a digital signal processor (DSP) that collects, collates, processes, and filters the digitized data set. The processor is also responsible for managing system timing and the imaging routines which can acquire real-time data at rates as high as 150 Hz. Many of the synchronously timed processes are controlled by a complex programmable logic device that is also used in conjunction with the DSP to orchestrate data flow. The operation of the system is implemented through a comprehensive graphical user interface designed with LABVIEW software which integrates automated calibration, data acquisition, data organization, and signal postprocessing. Performance analysis demonstrates very low system noise (approximately 1 pW rms noise equivalent power), excellent signal precision (<0.04%-0.2%) and long term system stability (<1% over 40 min). A large dynamic range (approximately 190 dB) accommodates a wide scope of measurement geometries and tissue types. First experiments on tissue phantoms show that dynamic behavior is accurately captured and spatial location can be correctly tracked using this system.     

Dynamic behavior of amplitude detection Kelvin force microscopy in ultrahigh vacuum

The acquisition rate of all scanning probe imaging techniques with feedback control is limited by the dynamic response of the control loops. Performance criteria are the control loop bandwidth and the output signal noise power spectral density. Depending on the acceptable noise level, it may be necessary to reduce the sampling frequency below the bandwidth of the control loop. In this work, the frequency response of a vacuum Kelvin force microscope with amplitude detection (AM-KFM) using a digital signal processing (DSP) controller is characterized and optimized. Then, the main noise source and its impact on the output signal is identified. A discussion follows on how the system design can be optimized with respect to output noise. Furthermore, the interaction between Kelvin and distance control loop is studied, confirming the beneficial effect of KFM on topography artefact reduction in the frequency domain. The experimental procedure described here can be generalized to other systems and allows to locate the performance limitations.     

IEWT和FSK在齿轮与滚动轴承故障诊断中的应用

改进的经验小波变换方法(improved empirical wavelet transform,简称IEWT)是一种新的自适应性信号处理方法,将这种方法和快速谱峭度(fast spectral kurtosis,简称FSK)相结合,进行齿轮与滚动轴承的故障诊断。首先,采用IEWT对信号进行分解,筛选出故障特征最为明显的2个分量并重构信号;其次,对重构信号进行快速谱峭度滤波;最后,对滤波后的信号进行包络谱分析,提取出信号的故障特征。分析齿轮断齿及滚动轴承故障信号,与直接包络谱和基于EMD经验模态分解(empirical mode decomposition,简称EMD)方法的FSK滤波包络谱分析方法相比可知,采用IEWT处理后再进行FSK滤波的信号进行包络谱分析更具有区分性,可有效识别齿轮和滚动轴承的故障特征。     

用于宽带光谱滤波的光学衍射神经网络的设计

光谱处理在光学研究和应用中具有重要意义。针对特定任务,已经开发了各种设备和仪器进行光谱的滤波、整形与分析、波长解复用等,但还没有具有先进处理能力的多任务光谱处理设备。设计了一种用于光谱滤波的衍射神经网络,其由相位调制型衍射层与探测层构成。在训练过程中加入了波长参数,以实现对宽带信号的处理;通过损失函数的设计,可以对输出光谱进行控制。以可见光波段的宽带信号为例,实现了单、双通带光谱滤波,且中心波段的宽度和相对强度可调节。证明了该光学衍射神经网络可以有效处理宽带光谱,并为实现更复杂的光谱处理任务奠定了基础。     

Precision of light intensity measurement in biological optical microscopy

Standardization and calibration of optical microscopy systems have become an important issue owing to the increasing role of biological imaging in high‐content screening technology. The proper interpretation of data from high‐content screening imaging experiments requires detailed information about the capabilities of the systems, including their available dynamic range, sensitivity and noise. Currently available techniques for calibration and standardization of digital microscopes commonly used in cell biology laboratories provide an estimation of stability and measurement precision (noise) of an imaging system at a single level of signal intensity. In addition, only the total noise level, not its characteristics (spectrum), is measured. We propose a novel technique for estimation of temporal variability of signal and noise in microscopic imaging. The method requires registration of a time series of images of any stationary biological specimen. The subsequent analysis involves a multi‐step process, which separates monotonic, periodic and random components of every pixel intensity change in time. The technique allows simultaneous determination of dark, photonic and multiplicative components of noise present in biological measurements. Consequently, a respective confidence interval (noise level) is obtained for each level of signal. The technique is validated using test sets of biological images with known signal and noise characteristics. The method is also applied to assess uncertainty of measurement obtained with two CCD cameras in a wide‐field microscope.     

多道成像光谱探测列阵的研究

本文介绍一种多道成象光谱探测列阵的工作原理及设计思想。通过对器件主要技术参数的分析及低功耗扫描电路的应用,并采取场注入等平面工艺措施,使探测列阵的光敏象元结构得到改善,从而提高了探测器的响应度和动态范围,光谱分析范围则可由近红外区到紫外区。最后给出主要参数的实验数据。     

Spectral resolution in conjunction with polar plots improves the accuracy and reliability of FLIM measurements and estimates of FRET efficiency

A spectrograph with continuous wavelength resolution has been integrated into a frequency‐domain fluorescence lifetime‐resolved imaging microscope (FLIM). The spectral information assists in the separation of multiple lifetime components, and helps resolve signal cross‐talking that can interfere with an accurate analysis of multiple lifetime processes. This extends the number of different dyes that can be measured simultaneously in a FLIM measurement. Spectrally resolved FLIM (spectral‐FLIM) also provides a means to measure more accurately the lifetime of a dim fluorescence component (as low as 2% of the total intensity) in the presence of another fluorescence component with a much higher intensity. A more reliable separation of the donor and acceptor fluorescence signals are possible for Förster resonance energy transfer (FRET) measurements; this allows more accurate determinations of both donor and acceptor lifetimes. By combining the polar plot analysis with spectral‐FLIM data, the spectral dispersion of the acceptor signal can be used to derive the donor lifetime – and thereby the FRET efficiency – without iterative fitting. The lifetime relation between the donor and acceptor, in conjunction with spectral dispersion, is also used to separate the FRET pair signals from the donor alone signal. This method can be applied further to quantify the signals from separate FRET pairs, and provide information on the dynamics of the FRET pair between different states.     

Full spectrum filterless fluorescence microscopy

In conventional microscopes, fluorescence emission is separated from the backscattered illumination using the Stokes shift, whereby the emission occurs at a longer wavelength to the excitation. Such separation is usually achieved through a combination of wavelength filters that divide the spectrum into mutually exclusive excitation and emission bands. It is therefore impossible in these microscopes to access the full excitation/emission spectrum of the specimen in a single image. We report on a microscope that acquired fluorescence images using illumination across the spectral range 450–680 nm; the full emission spectrum was detected simultaneously across the same range. The microscope was also combined with structured illumination optical sectioning to give three-dimensionally resolved images with improved background rejection. Full spectrum fluorescence images of biological specimens are demonstrated. As this system is more versatile than the standard fluorescence microscope, it could be of benefit in many fluorescence imaging applications.     

利用光谱滤波技术提高电视跟踪系统作用距离的研究

详细分析了目标、天空背景的光谱辐射特性、大气条件以及电视跟踪系统各环节对探测靶面上目标与背景对比度的影响,确定了利用光谱滤波提高电视跟踪系统作用距离的理论基础。实测了作为跟踪目标的3号曳光弹的光谱辐射特性;根据与电视跟踪系统工作环境相近的大气模式,从理论上计算、分析了天空背景的光谱辐射特性。通过计算机模拟与实测各种滤光片对天空背景辐射的抑制效果及对模拟目标信号衰减程度相结合的办法,从理论及实验上研究了光谱滤波提高电视跟踪系统作用距离的规律。并且从光谱滤波角度提出了对系统所采用的探测器及合作目标信号的某些特性的改进意见。对九种不同背景条件下光谱滤波效果的计算机模拟分析表明:光谱滤波能稳定有效地提高探测器靶面上的目标与背景的对比度。对3号曳光弹,在海平面45km,水平路径上.在0.810μm,处的窄带光谱滤波能使探测器靶面上的对比度提高4～7倍,作用距离可由40km提高到(50～53)km。实验表明:光谱滤波后,本征对比度可提高1.25～3.59倍。     

医用显微成像光谱仪的光谱定标技术

为了能对自主研制的脑肿瘤手术医用显微成像光谱仪进行光谱定标,设计了由单色仪、钨灯光源、棱镜-光栅-棱镜成像光谱仪及手术显微平台组成的光谱定标系统。采用单色仪波长扫描法,自主开发了相应的光谱定标系统软件,获得了显微成像光谱仪全谱段的光谱数据,完成了数据处理和分析等工作。通过调整光路、单色仪定标、成像光谱仪定标3个步骤实现了系统的光谱定标。定标结果表明:显微成像光谱仪的光谱区大于400~900nm;定标精度高于0.1nm,光谱分辨率高于3nm,各项特征指标均高于设计指标。测试验证实验表明,所建立的光谱定标系统定标精准,结构简单、紧凑,操作简单,符合显微成像光谱仪的实际临床应用要求。     

The use of low-cost frame grabber boards on PC-AT computers for ESI: possibilities and limitations

A Zeiss EM902 microscope was connected to a PC-AT computer, equipped with a Data Translation DT2861 frame grabber board. Using this system, the three-window background extrapolation algorithm was implemented to perform elemental mapping in the electron spectroscopic imaging mode of an energy filtering transmission electron microscope.