Direct electronic visualization of high-voltage TEM images by sequential pixel acquisition

A hybrid imaging mode is described that allows direct electronic detection and manipulation of TEM images. Such imaging mode should be of particular interest and usefulness in high-voltage instruments, in which image visibility is notably poor, but it can also be adapted to conventional electron microscopes. Pixels are detected sequentially by electron-optically sweeping the whole transmitted beam in a two-dimensional raster past a defining aperture located below the camera. Pixel data are obtained directly in digital form and can thus be conveniently manipulated on-line or stored for subsequent processing. Preliminary results obtained in one dimension confirm satisfactorily the anticipated characteristics of two-dimensional images. Possible consequences, applications and extensions of this electronic image acquisition mode are discussed.     

Three-dimensional image formation in confocal microscopy

Three-dimensional (3-D) imaging in confocal microscopes is considered in terms of 3-D transfer functions. This leads to an explanation of axial imaging properties. The axial response was observed in both object-scanning and beam-scanning microscopes and the influence of off-axis examination investigated. By simple processing of multi-detector signals, imaging in both the axial and transverse directions can be improved.     

Maximum pixel spectrum: a new tool for detecting and recovering rare, unanticipated features from spectrum image data cubes

A new software tool, the maximum pixel spectrum, detects rare events within a spectrum image data cube, such as that generated with electron‐excited energy‐dispersive X‐ray spectrometry in a scanning electron microscope. The maximum pixel spectrum is a member of a class of ‘derived spectra’ that are constructed from the spectrum image data cube. Similar to a conventional spectrum, a derived spectrum is a linear array of intensity vs. channel index that corresponds to photon energy. A derived spectrum has the principal characteristics of a real spectrum so that X‐ray peaks can be recognized. A common example of a derived spectrum is the summation spectrum, which is a linear array in which the summation of all pixels within each energy plane gives the intensity value for that channel. The summation spectrum is sensitive to the dominant features of the data cube. The maximum pixel spectrum is constructed by selecting the maximum pixel value within each X‐ray energy plane, ignoring the remaining pixels. Peaks corresponding to highly localized trace constituents or foreign contaminants, even those that are confined to one pixel of the image, can be seen at a glance when the maximum pixel spectrum is compared with the summation spectrum.     

An electron beam lithography and digital image acquisition system for scanning electron microscopes

A low‐cost microcontroller based control and data acquisition unit for digital image recording of scanning electron microscope (SEM) images and scanning electron microscope based electron beam lithography (EBL) is described. The developed microcontroller low‐level embedded software incorporates major time critical functions for image acquisition and electron beam lithography and makes the unit an intelligent module which communicates via USB with the main computer. The system allows recording of images with up to 4096 × 4096 pixel size, different scan modes, controllable dwell time, synchronization with main power frequency, and other user controllable functions. The electron beam can be arbitrary positioned with 12‐bit precision in both dimensions and this is used to extend the scanning electron microscope capabilities for electron beam lithography. Hardware and software details of the system are given to allow its easy duplication. Performance of the system is discussed and exemplary results are presented.     

Reduction in acquisition time of scanning electron microscopy image using complex hysteresis smoothing

Complex hysteresis smoothing (CHS), which was developed for noise removal of scanning electron microscopy (SEM) images some years ago, is utilized in acquisition of an SEM image. When using CHS together, recording time can be reduced without problems by about one-third under the condition of SEM signal with a comparatively high signal-to-noise ratio (SNR). We do not recognize artificiality in a CHS-filtered image, because it has some advantages, that is, no degradation of resolution, only one easily chosen processing parameter (this parameter can be fixed and used in this study), and no processing artifacts. This originates in the fact that its criterion for distinguishing noise depends simply on the amplitude of the SEM signal. The automation of reduction in acquisition time is not difficult, because CHS successfully works for almost all varieties of SEM images with a fairly high SNR.     

A method for personal expertise-independent evaluation of image resolution in scanning electron microscopy

The two conventional methods currently employed for the evaluation of image resolution in scanning electron microscopy are the gap method and a fast Fourier transform (FFT) method. These can be highly dependent on personal expertise on the distinction between signal information and noise contained in a micrograph. Hence, the present paper proposes an alternative method (referred to as a contrast-to-gradient (CG) method) that can determine the image resolution of a micrograph without requiring personal expertise on the judgment of noise. The image resolution in the CG method is defined as a weighted harmonic mean of the local resolution, which is proportional to the quotient of the threshold contrast divided by the local gradient. The local gradient is calculated from the quadratic function that best fits the local pixel intensities over 5 x 5 pixels. It has been shown that the CG method, compared with the FFT method, has a broader range of applications for various types of images, such as low-contrast, noise-containing, filter-processed, highly directional, and quasi-periodic feature images.     

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.     

A nondestructive method for three-dimensional reconstruction of luminescence materials: Principles,data acquisition,image processing

In the study presented here we have tried to state the principles and calculate and visualize models of three-dimensional (3-D)-cathodoluminescence reconstruction of luminescence structures by scanning electron microscopy (SEM). The new technique does not destroy the specimen and uses the variable energy of the electron beam to penetrate to different depths in the specimen volume. The SEM in color cathodoluminescence mode (CCL-SEM) detects integrated panchromatic CL-images for different energies of the electron beam. The use of electron scattering theory in solids and theories of cathodoluminescence and color allow the production of problem-oriented software for the routine processing of primary images. Processed images represent the CCL-SEM displays of separated layers (without CL information from other layers) up to the maximum depth penetrated by the beam. The 3-D reconstruction is carried out through algorithms developed using a personal computer, software, and a set of processed two-dimensional (2-D) images. The first experimental work was accomplished using a multilayer SiC mesastructure. The final reconstructed image of SiC material demonstrates separated epitaxial layers of different SiC polytypes and Z sections (YOZ and XOZ sections). The 3-D image represents the space distribution of CL-spectral data in color CL interpretation.     

New developments in fast image processing and data acquisition for STM

We have developed new easy-to-use but powerful software in order to obtain the best scan rates in data acquisition and image processing for a PC-AT microcomputer. The programs have been completely written in the C-language, which made it possible to combine low-level characteristics (quick access to memory, interrupt control, etc.) without losing the advantages of a high-level language. The data acquisition software was intended to be sufficiently flexible to acquire either topographical or spectroscopic data with no configuration changes. In topographical (z=f(x,y)) and spectroscopic (I=f(V, z)) measurements, we can obtain up to 15,000 samples/s rates, with 12-bit resolution in a ±10V range. Spectroscopic measurements are done by acquiring I(V) with the feedback off at several z values (up to 256) until I reaches a preset value; afterwards, z is reset automatically to the initial voltage to avoid tip-sample contact. The image processing was designed to get the best performances in time and quality with a simple system such as a PC-AT equipped with an IBM Professional Graphics Display (640times480 pixels and 256 simultaneous colours). It was structured in three parts: plane subtraction, image display (3-D shaded surfaces, 3-D surfaces with colour scales or 2-D contour maps), and filtering (smoothing filters or FFT convolution-deconvolution techniques).     

TEM in materials science—past,present and future

The progress in transmission electron microscopy as applied in materials science is reviewed briefly, from the era of replica techniques in the 1940s and 1950s, through the development of the diffraction contrast technique in the late 1950s and 1960s, to the present day when instrumental resolution is sufficient to obtain structure images of a wide variety of crystalline solids. One of the most important advances has been the development of combined imaging and microanalytical techniques in a single instrument. The versatility of the TEM technique with atomic resolution and microanalytical facilities, the variety of signals and contrast effects available, and its universal application, establishes it as an essential tool in materials science for the foreseeable future.     

Porosity determination of ceramic materials by digital image analysis--a critical evaluation

Measuring the porosity of materials by digital image analysis of micrographs is a well-established and convenient method for the testing of metallic samples. However, when applied to ceramic materials, this method has been shown to be much less reliable and poorly reproducible. The purpose of this present work is to clarify the reason for this deficiency, involving many porosity measurements, performed on plasma-sprayed zirconia, under systematically varied microscopic imaging conditions, and the porosities being calculated using various evaluation methods. Comparison between of the results has shown that the present state of the image analysis method is not satisfactory for absolute porosity measurements on ceramic materials. It can be useful as a convenient tool for comparative measurements, however, if the imaging conditions maintained in the microscope and the evaluation method are held to be exactly identical.     

Dental gallium alloy composites studied by SEM and TEM

Analytical scanning and transmission electron microscopy (SEM and TEM) studies of dental gallium alloys have been carried out. The Ga alloys were made by triturating a LU powder (Ag–Sn–Cu rich alloy powder) and a GF powder (Ag–Sn–Cu–Pd rich alloy powder) with a liquid Ga alloy containing Ga, In and Sn. The dental materials were found to be composites consisting of remaining, undissolved particles from the Ag-based alloy powders in a matrix of reaction products with the Ga alloy. SEM studies have been carried out to give an overview of the composites. The distribution of the elements was found by the X-ray mapping technique. The phases in the matrix and the remaining alloy particles have been identified by electron diffraction, high-resolution electron microscopy and energy-dispersive X-ray spectroscopy. The following phases were identified in the LU alloy: orthorhombic Ag₃Sn, cubic Ag₉In₄, tetragonal β-Sn and hexagonal Ag₂Ga. In addition to these well-known phases Ga-rich regions were observed consisting of an intergrowth of tetragonal CuGa₂ and one of the cubic γ-Cu₉Ga₄ phases. In addition to these phases cubic Ga₇Pd₃ was found in the GF alloy. The anomalous setting expansion of the GF alloy may be explained by the presence of Ga₇Pd₃.     

Elimination of scanning electron microscopy image periodic distortions with digital signal-processing methods

Electromagnetic interference is one of the main distortion sources in scanning electron microscopy. Electromagnetic interference‐generated scanning electron microscopy image distortions are usually visible as edge blur (at low scan rates) or vibration (at high scan rates). Hardware solutions to this problem, e.g. electrostatic and magnetic shielding, are expensive and, in some cases, difficult to implement. The current investigations led to a significant decrease in the periodic distortions by a novel adaptation of software‐based digital signal processing to scanning electron microscopy problems, without any hardware modification.     

Photon event distribution sampling: an image formation technique for scanning microscopes that permits tracking of sub-diffraction particles with high spatial and temporal resolutions

In photon event distribution sampling, an image formation technique for scanning microscopes, the maximum likelihood position of origin of each detected photon is acquired as a data set rather than binning photons in pixels. Subsequently, an intensity-related probability density function describing the uncertainty associated with the photon position measurement is applied to each position and individual photon intensity distributions are summed to form an image. Compared to pixel-based images, photon event distribution sampling images exhibit increased signal-to-noise and comparable spatial resolution. Photon event distribution sampling is superior to pixel-based image formation in recognizing the presence of structured (non-random) photon distributions at low photon counts and permits use of non-raster scanning patterns. A photon event distribution sampling based method for localizing single particles derived from a multi-variate normal distribution is more precise than statistical (Gaussian) fitting to pixel-based images. Using the multi-variate normal distribution method, non-raster scanning and a typical confocal microscope, localizations with 8 nm precision were achieved at 10 ms sampling rates with acquisition of ~200 photons per frame. Single nanometre precision was obtained with a greater number of photons per frame. In summary, photon event distribution sampling provides an efficient way to form images when low numbers of photons are involved and permits particle tracking with confocal point-scanning microscopes with nanometre precision deep within specimens.     

航空光电成像电子稳像技术

介绍了一种电子稳像技术,解决了航空平台光电成像系统视频图像帧内运动模糊和帧间不稳定问题。通过建立运动模糊的数学模型,构建二维运动模糊点扩散函数,采用维纳滤波方法,消除图像帧内运动模糊;通过灰度投影算法,检测序列图像当前帧和参考帧之间的运动矢量,采用图像补偿方法,实现序列图像稳定输出。实验结果表明,本文方法能提高信噪比,实现1 pixel的稳定精度,有效改善图像质量,输出清晰稳定的视频图像, 具有精确、快速、实用的特点。     

Digital image acquisition in in vivo confocal microscopy

A flexible system for the real-time acquisition of in vivo images has been developed. Images are generated using a tandem scanning confocal microscope interfaced to a low-light-level camera. The video signal from the camera is digitized and stored using a Gould image processing system with a real-time digital disk (RTDD). The RTDD can store up to 3200 512 times 512 pixel images at video rates (30 images s^?1). Images can be input directly from the camera during the study, or off-line from a Super VHS video recorder. Once a segment of experimental interest is digitized onto the RTDD, the user can interactively step through the images, average stable sequences, and identify candidates for further processing and analysis. Examples of how this system can be used to study the physiology of various organ systems in vivo are presented.     

Cation segregation in Nb16W18O94 using high angle annular dark field scanning transmission electron microscopy and image processing

We report the characterization of the complex oxide Nb₁₆W₁₈O₉₄ using high angle annular dark field imaging at 200 kV in a scanning transmission electron microscope. The results of this study suggest that the W and Nb cations are not uniformly distributed among the cation columns projected along [001] but that there is preferential segregation of the heavier species to certain column sites. In order to analyse the experimental data obtained, an image processing methodology has been developed which may also find application in locating specific motifs within a generally distorted image field.     

Primary considerations for image enhancement in high-pressure scanning electron microscopy

The mechanisms of electron beam scattering are examined to evaluate its effect on contrast and resolution in high-pressure scanning electron microscopy (SEM) techniques reported in the literature, such as moist-environment ambient-temperature SEM (MEATSEM) or environmental SEM (ESEM). The elastic and inelastic scattering cross-sections for nitrogen are calculated in the energy range 5–25 keV. The results for nitrogen are verified by measuring the ionization efficiency, and measurements are also made for water vapour. The effect of the scattered beam on the image contrast was assessed and checked experimentally for a step contrast function at 20 kV beam voltage. A considerable degree of beam scattering can be tolerated in high-pressure SEM operation without a significant degradation in resolution. The image formation and detection techniques in high-pressure SEM are considered in detail in the accompanying paper.     

Fractal compression and adaptive sampling: reducing the image acquisition time in scanning probe microscopy

In typical scanning probe microscope experiment a three‐dimensional image of a substrate is obtained. For a given scanning mechanism, the time needed to image an area depends mainly on the number of samples and the size of the image. The imaging speed is further compromised by drifts associated with the substrate and the piezoscanner. It is therefore desirable to improve the imaging speed with limited impact to the effective resolution of the resulting image. By utilizing an adaptive sampling scheme with fractal compression technique, we have demonstrated that the number of the required samples can be significantly reduced with minimal impact to the image quality. SCANNING 30: 463–473, 2008. © 2008 Wiley Periodicals, Inc.