Control of charging in low-voltage SEM

Charging of the specimen under electron beam irradiation is a common problem in scanning electron microscopy (SEM). It results in unstable imaging conditions and a loss in resolution due to defocus of the beam. In addition, it can cause permanent changes in some specimens from translocation of mobile ions under the influence of the induced electrostatic field. To minimize charging and its associated problems, the incident beam energy must be carefully chosen to be the value E₂ at which a dynamic charge balance is obtained. This article presents data on E₂ values for a variety of materials and demonstrates how E₂ is affected by the choice of angle of beam incidence.     

Fabrication and characterization of APT specimens from high dose heavy ion irradiated materials

The next generations of advanced energy systems will require materials that can withstand high doses of irradiation at elevated temperatures. Therefore, a methodology has been developed for the fabrication of high-dose ion-irradiated atom probe tomography specimens at a specific dose with the use of a focused ion beam milling system. The method also enables the precise ion dose of the atom probe tomography specimen to be estimated from the local concentration of the implanted ions. The method has been successfully applied to the characterization of the distribution of nanoclusters in a radiation-tolerant 14YWT nanostructured ferritic steel under ion irradiation to doses up to 400 displacements per atom.     

Charging effect in electron-irradiated ice

To maintain the original distribution pattern of diffusible elements in biological samples, electron probe microanalysis is carried out with frozen hydrated bulk specimens and cryosections, analysed at temperatures below 130 K. Ice has a very low intrinsic conductivity at this working temperature and surface- and space-charging appears, when uncoated specimens are irradiated with non-penetrating electrons. Although coating with a grounded conductor abolishes the surface potential, the build-up of an internal space-charge field is possible, depending on the sample thickness and beam voltage used. Consequently, the geometry of the X-ray source volume and the spectral distribution of the emitted continuous and characteristic X-rays are affected. To simulate the situation for microanalysis of frozen hydrated specimens the charging process in electron irradiated ice is studied by recording simultaneous specimen currents from the top and bottom of ice layer preparations. The external currents yield information on the build-up of internal space-charge fields which result from the balance of charge injection, storage, and transport. Irradiation of uncoated bulk specimens with a finely focused beam results in the build-up of a space-charge field close to the surface, which causes a reduction of the depth of microprobe analysis. In coated bulk specimens the induced conductivity renders possible a current flow to the front electrode, thereby limiting the space-charge field. Sections with an effective rear electrode will not charge appreciably if the electron range is larger than about half the section thickness.     

A study of electron beam-induced conductivity in resists.

The charging of polymeric resist materials during electron beam irradiation leads to significant problems during imaging and lithography processes. Charging occurs because of charge deposition in the polymer and charge generation/trapping due to formation of electron-hole pairs in the dielectric. The presence of such charge also results in the phenomena of electron beam-induced conductivity (EBIC). Electron beam-induced conductivity data have been obtained for three commercial e-beam resists under a variety of dose rate and temperature conditions. From the observed values of induced conductivity under varying conditions significant information about the generation of electron-hole pair and the transport of charge in the resist can be obtained. Three electron beam resists, EBR900, ZEP7000, and PBS are examined by an external bias method. The difference in resist chemistry is considered to play the role in the initial state EBIC behaviors among three resists even though the way that it affects the behaviors is not clear. A comparison of the power consumption comparison is proposed as a measure to give a preliminary estimate of the carrier concentration and carrier drift velocity differences among the resists. A simple single trap model with constant activation energy is proposed and provides good agreement with experiment.     

Interpretation of secondary electron images obtained using a low vacuum SEM

Charging of insulators in a variable pressure environment was investigated in the context of secondary electron (SE) image formation. Sample charging and ionized gas molecules present in a low vacuum specimen chamber can give rise to SE image contrast. "Charge-induced" SE contrast reflects lateral variations in the charge state of a sample caused by electron irradiation during and prior to image acquisition. This contrast corresponds to SE emission current alterations produced by sub-surface charge deposited by the electron beam. "Ion-induced" contrast results from spatial inhomogeneities in the extent of SE signal inhibition caused by ions in the gaseous environment of a low vacuum scanning electron microscope (SEM). The inhomogeneities are caused by ion focusing onto regions of a sample that correspond to local minima in the magnitude of the surface potential (generated by sub-surface trapped charge), or topographic asperities. The two types of contrast exhibit characteristic dependencies on microscope operating parameters such as scan speed, beam current, gas pressure, detector bias and working distance. These dependencies, explained in terms of the behavior of the gaseous environment and sample charging, can serve as a basis for a correct interpretation of SE images obtained using a low vacuum SEM.     

Observation of the ion-mirror effect during microscopy of insulating materials

Utilizing the ion beam of a focused ion beam (FIB)/scanning electron microscope (SEM) microscope to investigate non‐conductive samples, we observe a mirror image very much similar to the one that is commonly obtained with the electron beam and the same samples. To our knowledge this is the first observation of what can be called ‘Ion‐Mirror Effect’. This effect is produced by a positive charging of the sample obtained by rastering with high‐energy ions (30 kV) and a subsequent imaging with low energy ones (5 kV). The proposed explanation is that first a positive charge is trapped within the sample and eventually the lower energy ions are deflected back by the latter, and hit the surface of the microscope chamber very similar to what happens in the ‘Electron‐Mirror Effect’. The mirror image is produced after detection of the electrons produced by the interaction between ions and the chamber materials.     

Automated in‐chamber specimen coating for serial block‐face electron microscopy

When imaging insulating specimens in a scanning electron microscope, negative charge accumulates locally (‘sample charging’). The resulting electric fields distort signal amplitude, focus and image geometry, which can be avoided by coating the specimen with a conductive film prior to introducing it into the microscope chamber. This, however, is incompatible with serial block‐face electron microscopy (SBEM), where imaging and surface removal cycles (by diamond knife or focused ion beam) alternate, with the sample remaining in place. Here we show that coating the sample after each cutting cycle with a 1–2 nm metallic film, using an electron beam evaporator that is integrated into the microscope chamber, eliminates charging effects for both backscattered (BSE) and secondary electron (SE) imaging. The reduction in signal‐to‐noise ratio (SNR) caused by the film is smaller than that caused by the widely used low‐vacuum method. Sample surfaces as large as 12 mm across were coated and imaged without charging effects at beam currents as high as 25 nA. The coatings also enabled the use of beam deceleration for non‐conducting samples, leading to substantial SNR gains for BSE contrast. We modified and automated the evaporator to enable the acquisition of SBEM stacks, and demonstrated the acquisition of stacks of over 1000 successive cut/coat/image cycles and of stacks using beam deceleration or SE contrast.     

In-flight calibration of mesospheric rocket plasma probes

Many effects and factors can influence the efficiency of a rocket plasma probe. These include payload charging, solar illumination, rocket payload orientation and rotation, and dust impact induced secondary charge production. As a consequence, considerable uncertainties can arise in the determination of the effective cross sections of plasma probes and measured electron and ion densities. We present a new method for calibrating mesospheric rocket plasma probes and obtaining reliable measurements of plasma densities. This method can be used if a payload also carries a probe for measuring the dust charge density. It is based on that a dust probe's effective cross section for measuring the charged component of dust normally is nearly equal to its geometric cross section, and it involves the comparison of variations in the dust charge density measured with the dust detector to the corresponding current variations measured with the electron and/or ion probes. In cases in which the dust charge density is significantly smaller than the electron density, the relation between plasma and dust charge density variations can be simplified and used to infer the effective cross sections of the plasma probes. We illustrate the utility of the method by analysing the data from a specific rocket flight of a payload containing both dust and electron probes.     

Dynamic secondary electron contrast effects in liquid systems studied by environmental scanning electron microscopy

We report an investigation into a dynamic contrast phenomenon in water-oil emulsions imaged in the environmental scanning electron microscope. Secondary electron contrast between oil and water phases is shown to change with scan rate, even inverting in extreme cases. This effect is attributed to the fact that charge carriers in liquids have intermediate mobilities compared with those in metallic conductors and solid insulators. Thus, increasing the electron energy flux density (via slower scan rates) results in the temporary accumulation of excess charge, which in turn gives rise to increased secondary electron emission. Excess charge dissipates between frames, however, such that classical charging of the specimen is not observed. The oils used here have conductivities lower than that of water, making them more susceptible to the effect. However, the material within the primary electron interaction volume is a conductive medium. We demonstrate that charging effects are not seen in regions of the oil where the interaction volume is in contact with the more conductive continuous water phase. Secondary electron emission from these regions therefore approximates to the intrinsic yield.     

Bias-induced forces in conducting atomic force microscopy and contact charging of organic monolayers

Contact electrification, a surface property of bulk dielectric materials, has now been observed at the molecular scale using conducting atomic force microscopy (AFM). Conducting AFM measures the electrical properties of an organic film sandwiched between a conducting probe and a conducting substrate. This paper describes physical changes in the film caused by the application of a bias. Contact of the probe leads to direct mechanical stress and the applied electric field results in both Maxwell stresses and electrostriction. Additional forces arise from charge injection (contact charging). Electrostriction and contact charging act oppositely from the normal long-range Coulomb attraction and dominate when a charged tip touches an insulating film, causing the tip to deflect away from the film at high bias. A bias-induced repulsion observed in spin-coated PMMA films may be accounted for by either mechanism. In self-assembled monolayers, however, tunnel current signals show that the repulsion is dominated by contact charging.     

Quantitative measurements of charging in a gaseous environment

Charge accumulation in insulating or semiconducting samples due to electron beam irradiation is one of the key problems in electron microscopy. One of the most promising techniques for reducing the severity of such charging is to surround the sample with a low-pressure atmosphere of a gas. The charging behavior of a number of materials, surrounded by a variety of gases, has been determined to identify the important factors which control charging under these conditions. The magnitude of the surface potential was deduced from an analysis of x-ray spectra from the surface. The relationship between surface charge, gas pressure, and gas type are measured, and the charging reduction efficiency (CRE) is compared.     

High-pressure scanning electron microscopy of insulating materials: A new approach

A new SEM technique for imaging uncoated non-conducting specimens at high beam voltages is described which employs a high-pressure environment and an electric field to achieve charge neutralization. During imaging, the specimen surface is kept at a stable low voltage, near earth potential, by directing a flow of positive gas ions at the specimen surface under the action of an electric bias field at a pressure of about 200 Pa. In this way charge neutrality is continuously maintained to obtain micrographs free of charging artefacts. Images are formed by specimen current detection containing both secondary electron and backscattered electron signal information. Micrographs of geological, ceramic, and semiconductor materials obtained with this method are presented. The technique is also useful for the SEM examination of histological sections of biological specimens without any further preparation. A simple theory for the charge neutralization process is described. It is based on the interaction of the primary and emissive signal components with the surrounding gas medium and the resulting neutralizing currents. Further micrographs are presented to illustrate the pressure dependence of the charge neutralization process in two glass specimens which show clearly identifiable charging artefacts in conventional microscopy.     

Reduction of charging in protein electron cryomicroscopy

Charging causes a loss of resolution in electron cryomicroscopy with biological specimens prepared without a continuous carbon support film. Thin conductive films were deposited onto catalase crystals prepared across holes using ion-beam sputtering and thermal evaporation and evaluated for the effectiveness of charge reduction. Deposits applied by ion-beam sputtering reduced charging but concurrently resulted in structural damage. Coatings applied by thermal evaporation also reduced charging, and preserved the specimen structure beyond 5 Å resolution as judged from electron diffraction patterns and images of glucose-embedded catalase crystals tilted to 45° in the microscope. This study demonstrates for the first time the feasibility of obtaining high-resolution data from unstained, unsupported protein crystals with a conductive surface coating.     

High‐performance serial block‐face SEM of nonconductive biological samples enabled by focal gas injection‐based charge compensation

A longstanding limitation of imaging with serial block‐face scanning electron microscopy is specimen surface charging. This charging is largely due to the difficulties in making biological specimens and the resins in which they are embedded sufficiently conductive. Local accumulation of charge on the specimen surface can result in poor image quality and distortions. Even minor charging can lead to misalignments between sequential images of the block‐face due to image jitter. Typically, variable‐pressure SEM is used to reduce specimen charging, but this results in a significant reduction to spatial resolution, signal‐to‐noise ratio and overall image quality. Here we show the development and application of a simple system that effectively mitigates specimen charging by using focal gas injection of nitrogen over the sample block‐face during imaging. A standard gas injection valve is paired with a precisely positioned but retractable application nozzle, which is mechanically coupled to the reciprocating action of the serial block‐face ultramicrotome. This system enables the application of nitrogen gas precisely over the block‐face during imaging while allowing the specimen chamber to be maintained under high vacuum to maximise achievable SEM image resolution. The action of the ultramicrotome drives the nozzle retraction, automatically moving it away from the specimen area during the cutting cycle of the knife. The device described was added to a Gatan 3View system with minimal modifications, allowing high‐resolution block‐face imaging of even the most charge prone of epoxy‐embedded biological samples.     

Broadening the applications of the atom probe technique by ultraviolet femtosecond laser

Laser assisted field evaporation using ultraviolet (UV) wavelength gives rise to better mass resolution and signal-to-noise ratio in atom probe mass spectra of metals, semiconductors and insulators compared to infrared and green lasers. Combined with the site specific specimen preparation techniques using the lift-out and annular Ga ion milling in a focused ion beam machine, a wide variety of materials including insulating oxides can be quantitatively analyzed by the three-dimensional atom probe using UV laser assisted field evaporation. After discussing laser irradiation conditions for optimized atom probe analyses, recent atom probe tomography results on oxides, semiconductor devices and grain boundaries of sintered magnets are presented.     

氦离子显微镜：技术和应用进展

氦离子显微镜能够提供所有扫描射线法中最高空间分辨率的表面图像,以及极高的表面灵敏度。这些因素结合起来,能提供纳米级的无与伦比的样品信息。结合这种工具独特的充电控制技术,一些应用问题可以得到解决。本文介绍了一些近期的结果。例如,生命科学影像往往依赖于研究高度绝缘的轻型有机材料的表面形貌。氦离子显微镜无需导电涂层就可以提供极好的成像。诸如生物工程学研究之类的应用,可同时受益于充电控制和氦离子成像可避免电子束辐射造成损伤的优点。可以使用氦离子显微镜进行离子散射能谱分析,从而将这项灵敏度最高的表面技术应用于同时要求具有空间分辨率的分析任务中。氦离子显微镜也用于表面和材料的图形化。这对于创建和表征纳米级的新特性是至关重要的。     

Liquid metal alloy ion source based metal ion injection into a room-temperature electron beam ion source

We have carried out a series of measurements demonstrating the feasibility of using the Dresden electron beam ion source (EBIS)-A, a table-top sized, permanent magnet technology based electron beam ion source, as a charge breeder. Low charged gold ions from an AuGe liquid metal alloy ion source were injected into the EBIS and re-extracted as highly charged ions, thereby producing charge states as high as Au(60 +). The setup, the charge breeding technique, breeding efficiencies as well as acceptance and emittance studies are presented.     

Operation of the CAPRICE electron cyclotron resonance ion source applying frequency tuning and double frequency heating

The properties of the electromagnetic waves heating the electrons of the ECR ion sources (ECRIS) plasma affect the features of the extracted ion beams such as the emittance, the shape, and the current, in particular for higher charge states. The electron heating methods such as the frequency tuning effect and the double frequency heating are widely used for enhancing the performances of ECRIS or even for the routine operation during the beam production. In order to better investigate these effects the CAPRICE ECRIS has been operated using these techniques. The ion beam properties for highly charged ions have been measured with beam diagnostic tools. The reason of the observed variations of this performance can be related to the different electromagnetic field patterns, which are changing inside the plasma chamber when the frequency is varying.     

Generation of high charge state metal ion beams by electron cyclotron resonance heating of vacuum arc plasma in cusp trap

A method for generating high charge state heavy metal ion beams based on high power microwave heating of vacuum arc plasma confined in a magnetic trap under electron cyclotron resonance conditions has been developed. A feature of the work described here is the use of a cusp magnetic field with inherent "minimum-B" structure as the confinement geometry, as opposed to a simple mirror device as we have reported on previously. The cusp configuration has been successfully used for microwave heating of gas discharge plasma and extraction from the plasma of highly charged, high current, gaseous ion beams. Now we use the trap for heavy metal ion beam generation. Two different approaches were used for injecting the vacuum arc metal plasma into the trap--axial injection from a miniature arc source located on-axis near the microwave window, and radial injection from sources mounted radially at the midplane of the trap. Here, we describe preliminary results of heating vacuum arc plasma in a cusp magnetic trap by pulsed (400 μs) high power (up to 100 kW) microwave radiation at 37.5 GHz for the generation of highly charged heavy metal ion beams.     

Adhesion of microbes using 3-aminopropyl triethoxy silane and specimen stabilisation techniques for analytical transmission electron microscopy

A variety of adhesive support-films were tested for their ability to adhere various biological specimens for transmission electron microscopy. Support films primed with 3-amino-propyl triethoxy silane (APTES), poly- l -lysine, carbon and ultraviolet-B (UV-B)-irradiated carbon were tested for their ability to adhere a variety of biological specimens including axenic cultures of Bacillus subtilis and Escherichia coli and wild-type magnetotactic bacteria. The effects of UV-B irradiation on the support film in the presence of air and electrostatic charge on primer deposition were tested and the stability of adhered specimens on various surfaces was also compared. APTES-primed UV-B-irradiated Pioloform^TM was consistently the best adhesive, especially for large cells, and when adhered specimens were UV-B irradiated they became remarkably stable under an electron beam. This assisted the acquisition of in situ phase-contrast lattice images from a variety of biominerals in magnetotactic bacteria, in particular metastable greigite magnetosomes. Washing tests indicated that specimens adhering to APTES-primed UV-B-irradiated Pioloform^TM were covalently coupled. The electron beam stability was hypothesised to be the result of mechanical strengthening of the specimen and support film and the reduced electrical resistance in the specimen and support film due to their polymerization and covalent coupling.