Cryogenic‐temperature electron microscopy direct imaging of carbon nanotubes and graphene solutions in superacids

Cryogenic electron microscopy (cryo‐EM) is a powerful tool for imaging liquid and semiliquid systems. While cryogenic transmission electron microscopy (cryo‐TEM) is a standard technique in many fields, cryogenic scanning electron microscopy (cryo‐SEM) is still not that widely used and is far less developed. The vast majority of systems under investigation by cryo‐EM involve either water or organic components. In this paper, we introduce the use of novel cryo‐TEM and cryo‐SEM specimen preparation and imaging methodologies, suitable for highly acidic and very reactive systems. Both preserve the native nanostructure in the system, while not harming the expensive equipment or the user. We present examples of direct imaging of single‐walled, multiwalled carbon nanotubes and graphene, dissolved in chlorosulfonic acid and oleum. Moreover, we demonstrate the ability of these new cryo‐TEM and cryo‐SEM methodologies to follow phase transitions in carbon nanotube (CNT)/superacid systems, starting from dilute solutions up to the concentrated nematic liquid‐crystalline CNT phases, used as the ‘dope’ for all‐carbon‐fibre spinning. Originally developed for direct imaging of CNTs and graphene dissolution and self‐assembly in superacids, these methodologies can be implemented for a variety of highly acidic systems, paving a way for a new field of nonaqueous cryogenic electron microscopy.     

MFM and gas adsorption isotherm analysis of proton beam irradiated multi-walled carbon nanotubes

To enhance the gas adsorption properties and modify the physical properties of carbon nanotubes, multi-walled carbon nanotubes (MWCNTs) were irradiated by high-energy proton beams, and the physical properties including morphology and local surface structure were investigated by using a transmission electron microscope (TEM), magnetic force microscope (MFM) and a gas adsorption isotherm apparatus which can deeply probe the fine structure of surface. Interestingly, clearer MFM images were obtained from the proton irradiated samples which supports that carbon exhibits magnetism under proton bombardments, although the intrinsic magnetic property is not understood. The layering properties of argon on MWCNTs were measured from 59 to 69 K and the interaction of argon on the surface was analyzed. The calculated values of isosteric heat of adsorption demonstrated that higher interaction of gas molecules with surface is found from the proton irradiated MWCNTs. This result strongly supports that the local surface modification, partial defects, for example, were created due to the external high energy impacts. Our results are worthy to note that gas adsorption technique can provide the fine atomic resolution which beyond the one of TEM and MFM.     

10-kV diffractive imaging using newly developed electron diffraction microscope

A new electron diffraction microscope based on a conventional scanning electron microscope (SEM), for obtaining atomic-level resolution images without causing serious damage to the specimen, has been developed. This microscope in the relatively low-voltage region makes it possible to observe specimens at suitable resolution and record diffraction patterns. Using the microscope we accomplished 10-kV diffractive imaging with the iterative phase retrieval and reconstructed the structure of a multi-wall carbon nanotube with its finest feature corresponding to 0.34-nm carbon wall spacing. These results demonstrate the possibility of seamless connection between observing specimens by SEM and obtaining their images at high resolution by diffractive imaging.     

Scanning probe microscopy installed with nanotube probes and nanotube tweezers

We have developed well-controlled processes for the growth and manipulation of carbon nanotubes. The relatively thin multiwalled nanotubes were prepared with high purity by arc discharge with a high gas temperature. In the manipulation of nanotubes, the first crucial process is to prepare a nanotube array, so-called nanotube cartridge. We have found the alternated current electrophoresis of nanotubes by which nanotubes are aligned at the knife-edge of a disposal razor. The second important process is to transfer a nanotube from the nanotube cartridge onto a substrate in a scanning electron microscope. Using this method, we have developed nanotube probes and nanotube tweezers that operate in a scanning probe microscope (SPM). The nanotube probes have been applied for observation of biological samples and industrial samples to clarify their advantages. The nanotube tweezers have been demonstrated for their motion in scanning electron microscope and operated to carry a nanomaterial in a SPM.     

In situ measurements and transmission electron microscopy of carbon nanotube field-effect transistors

We present the design and operation of a transmission electron microscopy (TEM)-compatible carbon nanotube (CNT) field-effect transistor (FET). The device is configured with microfabricated slits, which allows direct observation of CNTs in a FET using TEM and measurement of electrical transport while inside the TEM. As demonstrations of the device architecture, two examples are presented. The first example is an in situ electrical transport measurement of a bundle of carbon nanotubes. The second example is a study of electron beam radiation effect on CNT bundles using a 200 keV electron beam. In situ electrical transport measurement during the beam irradiation shows a signature of wall- or tube-breakdown. Stepwise current drops were observed when a high intensity electron beam was used to cut individual CNT bundles in a device with multiple bundles.     

Damages of screen-printed carbon nanotube cold cathode during the field emission process

The field emission properties of the screen-printed carbon nanotube (CNT) composite cathode have close relationship with its microstructure. In this study, carbon nanotube composite cold cathode with ZnO nano-particles as binding material was prepared using screen-printing method. Electric field cycles were used to post-treat the carbon nanotube composite cold cathode. During the process of electric field cycle treatment, obvious heat-induced damages were observed from the cathode. Scanning electron microscope and transmission electron microscope were employed to analyze the morphology and microstructure of the cathode. The possible mechanisms responsible for damages were discussed.     

Direct electron imaging in electron microscopy with monolithic active pixel sensors

A new imaging device for dynamic electron microscopy is in great demand. The detector should provide the experimenter with images having sufficient spatial resolution at high speed. Immunity to radiation damage, accumulated during exposures, is critical. Photographic film, a traditional medium, is not adequate for studies that require large volumes of data or rapid recording and charge coupled device (CCD) cameras have limited resolution, due to phosphor screen coupling. CCD chips are not suitable for direct recording due to their extreme sensitivity to radiation damage. This paper discusses characterization of monolithic active pixel sensors (MAPS) in a scanning electron microscope (SEM) as well as in a transmission electron microscope (TEM). The tested devices were two versions of the MIMOSA V (MV) chip. This 1M pixel device features pixel size of 17 x 17 microm(2) and was designed in a 0.6 microm CMOS process. The active layer for detection is a thin (less than 20 microm) epitaxial layer, limiting the broadening of the electron beam. The first version of the detector was a standard imager with electronics, passivation and interconnection layers on top of the active region; the second one was bottom-thinned, reaching the epitaxial layer from the bottom. The electron energies used range from a few keV to 30 keV for SEM and from 40 to 400 keV for TEM. Deterioration of the image resolution due to backscattering was quantified for different energies and both detector versions.     

Transmission electron microscopy at 20 kV for imaging and spectroscopy

The electron optical performance of a transmission electron microscope (TEM) is characterized for direct spatial imaging and spectroscopy using electrons with energies as low as 20 keV. The highly stable instrument is equipped with an electrostatic monochromator and a C_S-corrector. At 20 kV it shows high image contrast even for single-layer graphene with a lattice transfer of 213 pm (tilted illumination). For 4 nm thick Si, the 200 reflections (271.5 pm) were directly transferred (axial illumination). We show at 20 kV that radiation-sensitive fullerenes (C₆₀) within a carbon nanotube container withstand an about two orders of magnitude higher electron dose than at 80 kV. In spectroscopy mode, the monochromated low-energy electron beam enables the acquisition of EELS spectra up to very high energy losses with exceptionally low background noise. Using Si and Ge, we show that 20 kV TEM allows the determination of dielectric properties and narrow band gaps, which were not accessible by TEM so far. These very first results demonstrate that low kV TEM is an exciting new tool for determination of structural and electronic properties of different types of nano-materials.     

Imaging Si nanoparticles embedded in SiO(2) layers by (S)TEM-EELS

Fabrication of systems in which Si nanoparticles are embedded in a thin silica layer is today mature for non-volatile memory and opto-electronics applications. The control of the different parameters (position, size and density) of the nanoparticles population is a key point to optimize the properties of such systems. A review of dedicated transmission electron microscopy (TEM) methods, which can be used to measure these parameters, is presented with an emphasis on those relying on electron energy-loss spectroscopy (EELS). Defocused bright-field imaging can be used in order to determine topographic information of a whole assembly of nanoparticles, but it is not efficient for looking at individual nanoparticles. High-resolution electron imaging or dark-field imaging can be of help in the case of crystalline particles but they always provide underestimated values of the nanocrystals population. EELS imaging in the low-energy-loss domain around the Si plasmon peak, which gives rise to strong signals, is the only way to visualize all Si nanoparticles within a silica film and to perform reliable size and density measurements. Two complementary types of experiments are investigated and discussed more extensively: direct imaging with a transmission electron microscope equipped with an imaging filter (EFTEM) and indirect imaging from spectrum-imaging data acquired with a scanning transmission electron microscope equipped with a spectrometer (STEM-PEELS). The direct image (EFTEM) and indirect set of spectra (STEM-PEELS) are processed in order to deliver images where the contribution of the silica matrix is minimized. The contrast of the resulting images can be enhanced with adapted numerical filters for further morphometric analysis. The two methods give equivalent results, with an easier access for EFTEM and the possibility of a more detailed study of the EELS signatures in the case of STEM-PEELS. Irradiation damage in such systems is also discussed.     

Behaviour of TEM metal grids during in-situ heating experiments

The stability of Ni, Cu, Mo and Au transmission electron microscope (TEM) grids coated with ultra-thin amorphous carbon (α-C) or silicon monoxide film is examined by in-situ heating up to a temperature in the range 500–850 °C in a transmission electron microscope. It is demonstrated that some grids can generate nano-particles either due to the surface diffusion of metal atoms on amorphous film or due to the metal evaporation/redeposition. The emergence of nano-particles can complicate experimental observations, particularly in in-situ heating studies of dynamic behaviours of nano-materials in TEM. The most widely used Cu grid covered with amorphous carbon is unstable, and numerous Cu nano-particles start to form once the heating temperature reaches 600 °C. In the case of Ni grid covered with α-C film, a large number of Ni nano-crystals occur immediately when the temperature approaches 600 °C, accompanied by the graphitization of amorphous carbon. In contrast, both Mo and Au grids covered with α-C film exhibit good stability at elevated temperature, for instance, up to 680 and 850 °C for Mo and Au, respectively, and any other metal nano-particles are detected. Cu grid covered Si monoxide thin film is stable up to 550 °C, but Si nano-crystals appear under intensive electron beam. The generated nano-particles are well characterized by spectroscopic techniques (EDXS/EELS) and high-resolution TEM. The mechanism of nano-particle formation is addressed based on the interactions between the metal grid and the amorphous carbon film and on the sublimation of metal.     

Multivariate statistical analysis of electron energy-loss spectroscopy in anisotropic materials

Recently, an expression has been developed to take into account the complex dependence of the fine structure in core-level electron energy-loss spectroscopy (EELS) in anisotropic materials on specimen orientation and spectral collection conditions [Y. Sun, J. Yuan, Phys. Rev. B 71 (2005) 125109]. One application of this expression is the development of a phenomenological theory of magic-angle electron energy-loss spectroscopy (MAEELS), which can be used to extract the isotropically averaged spectral information for materials with arbitrary anisotropy. Here we use this expression to extract not only the isotropically averaged spectral information, but also the anisotropic spectral components, without the restriction of MAEELS. The application is based on a multivariate statistical analysis of core-level EELS for anisotropic materials. To demonstrate the applicability of this approach, we have conducted a study on a set of carbon K-edge spectra of multi-wall carbon nanotube (MWCNT) acquired with energy-loss spectroscopic profiling (ELSP) technique and successfully extracted both the averaged and dichroic spectral components of the wrapped graphite-like sheets. Our result shows that this can be a practical alternative to MAEELS for the study of electronic structure of anisotropic materials, in particular for those nanostructures made of layered materials.     

IMPROVED FABRICATION METHOD FOR CARBON NANOTUBE PROBE OF ATOMIC FORCE MICROSCOPY（AFM）

An improved arc discharge method is developed to fabricate carbon nanotube probe of atomic force microscopy (AFM) here. First, silicon probe and carbon nanotube are manipulated under an optical microscope by two high precision microtranslators. When silicon probe and carbon nanotube are very close, several tens voltage is applied between them. And carbon nanotube is divided and attached to the end of silicon probe, which mainly due to the arc welding function. Comparing with the arc discharge method before, the new method here needs no coat silicon probe with metal film in advance, which can greatly reduce the fabrication's difficulty. The fabricated carbon nanotube probe shows good property of higher aspect ratio and can more accurately reflect the true topography of silicon grating than silicon probe. Under the same image drive force, carbon nanotube probe had less indentation depth on soft triblock copolymer sample than silicon probe. This showed that carbon nanotube probe has lower spring constant and less damage to the scan sample than silicon probe.     

Lateral force microscopy of multiwalled carbon nanotubes

Carbon nanotubes are usually imaged with the atomic force microscope (AFM) in non-contact mode. However, in many applications, such as mechanical manipulation or elasticity measurements, contact mode is used. The forces affecting the nanotube are then considerable and not fully understood. In this work lateral forces were measured during contact mode imaging with an AFM across a carbon nanotube. We found that, qualitatively, both magnitude and sign of the lateral forces to the AFM tip were independent of scan direction and can be concluded to arise from the tip slipping on the round edges of the nanotube. The dependence on the normal force applied to the tip and on the ratio between nanotube diameter and tip radius was studied. We show that for small values of this ratio, the lateral force signal can be explained with a simple geometrical model.     

Selective specimen preparation for TEM observation of the cross-section of individual carbon nanotube/metal junctions

We present here an efficient method to prepare a transmission electron microscopy (TEM) specimen for selective observation of the cross-section of individual nanoscale structures. As a typical example, the cross-sectional TEM observation of a quasi-one-dimensional material - a nano-electronic component based on an individual carbon nanotube - is presented.     

Double-layer coating for field-emission cryo-scanning electron microscopy—present state and applications

Imaging of fast-frozen samples is the most direct approach for electron microscopy of organic material. It prevents chemical fixation and drying artifacts. Frozen samples can be replicated and imaged in the transmission electron microscope (TEM), or they can be directly visualized in the cryo-scanning electron microscope (cryo-SEM). Double-layer coating combines these two techniques and many of their advantages. With this method, the frozen bulk sample is coated similar to the TEM-replica technique with, for example, a shadow of platinum (at an angle of 45°) and an additional layer of carbon. Then, the sample is cryo-transferred to an SEM equipped with a cold stage and imaged with the material-dependent backscattered electron signal that shows the platinum distribution. With this method, charging artifacts and the effects of beam damage are significantly reduced. Although currently the resolution of the replica technique cannot be surpassed, the method greatly facilitates the processing of brittle, rapidly frozen samples because no replica cleaning is necessary. This makes the method especially suitable for high-pressure frozen samples.     

Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and high-resolution scanning and scanning transmission electron microscopy

This paper describes the use of sputter coating to prepare detergent-extracted cytoskeletons for observation by scanning (SEM), scanning transmission (STEM), inverted contrast STEM, and transmission (TEM) electron microscopy. Sputtered coats of 1–2 nm of platinum or tungsten provide both an adequate secondary electron signal for SEM and good contrast for STEM and TEM. At the same time, the grain size of the coating is sufficiently fine to be just at (platinum) or below (tungsten) the limit of resolution for SEM and STEM. In TEM, the granular structure of platinum coats is resolved, and platinum decoration artifacts are observed on the surface of structures. The platinum is deposited as small islands with a periodic distribution that may reveal information about the underlying molecular structure. This method produces samples that are similar in appearance to replicas prepared by low-angle rotary shadowing with platinum and carbon. However, the sputter-coating method is easier to use; more widely available to investigators; and compatible with SEM, STEM, and TEM. It may also be combined with immunogold and other labeling methods. While TEM provides the highest resolution images of sputter-coated cytoskeletons, it also damages the specimens owing to heating in the beam. In SEM and STEM cytoskeletons are stable and the resolution is adequate to resolve individual microfilaments. The best single method for visualizing cytoskeletons is inverted contrast STEM, which images both the metal-coated cytoskeletal structures and electron-dense material within the nucleus and cytoplasm as white against a dark background. STEM and TEM were both suitable for visualizing colloidal gold particles in immunolabeled samples.     

Wear characteristics of diamond-coated atomic force microscope probe

In atomic force microscope (AFM) applications, the wear of the probe is undoubtedly a serious concern since it affects the integrity of the measurements. In this work, wear tests were performed using an AFM with lateral force monitoring capability with the aim to better understand the wear characteristics of diamond-coated probes. For the assessment of the probe wear, a transmission electron microscope (TEM) as well as a scanning electron microscope were utilized. The degree of the probe wear was quantified using the Archard's wear equation. The structure of the diamond-coated probe was analyzed by using the TEM and Raman spectroscopy. From the experimental results, two different wear characteristics, the gradual wear and the abrupt fracture of the diamond coating, were observed. In the case of gradual wear, the wear coefficient of the diamond-coated probe slid against a silicon nitride specimen was about 10(-5)-10(-6). It was also found that the wear rate significantly decreased with increase in the sliding distance. Raman spectroscopy analysis showed that the difference in the chemical structure of the diamond coating may induce the different wear phenomena. These results may be effectively utilized for fundamental understanding of nano-wear characteristics of AFM probes.     

Radiation damage in coronene, rubrene and p-terphenyl, measured for incident electrons of kinetic energy between 100 and 200 kev

We have measured the sensitivity of three highly conjugated organic compounds to electron irradiation. Using a 200 keV TEM, loss of crystallinity was determined from quantitative electron-diffraction measurements. Degradation of the molecular ring structure was monitored from fading of the 6 eV pi-excitation peak in the energy-loss spectrum. Measurements at incident energies between 30 keV and 100 eV were made using a scanning electron microscope (SEM), by recording gradual decay of the cathodoluminescence (CL) signal. Expressed in Grays, the energy dose required for CL decay in coronene is a factor of 30 lower than for destruction of crystallinity and a factor of 300 lower than for destruction of the molecular structure. Below 1 keV, the CL-decay cross section shows no evidence of a threshold effect, indicating that the damage involved is caused by valence-electron (rather than K-shell) excitation. Therefore even relatively radiation-resistant organic materials may undergo some form of damage when examined in a low-energy electron microscope or a low-voltage SEM.     

Electron holographic observation of micro-magnetic fields current-generated from single carbon coil

A carbon coil was evaluated for use as a micro-solenoid in a small magnetic device. A single carbon coil was lifted out of the aggregate using a tungsten fine probe in a focused ion beam (FIB) system and was wired to two small electrodes in the specimen holder of a transmission electron microscope (TEM). A direct current was supplied to the single carbon coil. A micro/nano-magnetic field generated from the coil was directly observed by electron holography. A computer simulation of electron holography was also done to quantitatively analyze the magnetic field. Details on the FIB technique, the electron holographic observation and the simulation are described.     

Fabrication of a versatile substrate for finding samples on the nanometer scale

With increasing interest in nanometer scale studies, a common research issue is the need to use different analytical systems with a universal substrate to relocate objects on the nanometer scale. Our paper addresses this need. Using the delicate milling capability of a focused ion beam (FIB) system, a region of interest (ROI) on a sample is labelled via a milled reference grid. FIB technology allows for milling and deposition of material at the sub 20-nm level, in a similar user environment as a standard scanning electron microscope (SEM). Presently commercially available transmission electron microscope (TEM) grids have spacings on the order 100 μm on average; this technique can extend this dimension down to the submicrometre level. With a grid on the order of a few micrometres optical, FIBs, TEMs, scanning electron microscopes (SEMs), and atomic force microscopes (AFM) are able to image the ROI, without special chemical processes or conductive coatings required. To demonstrate, Au nanoparticles of ∼ 25 nm in size were placed on a commercial Formvar^®- and carbon-coated TEM grid and later milled with a grid pattern. Demonstration of this technique is also extended to bulk glass substrates for the purpose of sample location. This process is explained and demonstrated using all of the aforementioned analytical techniques.