FAST-EM array tomography: a workflow for multibeam volume electron microscopy
Arent J. KievitsB. H. Peter DuinkerkenRyan LaneCecilia de HeusDaan van Beijeren Bergen en HenegouwenTibbe HöppenerAnouk H. G. WoltersNalan LivBen N. G. GiepmansJacob P. Hoogenboom
1
Citation
42
Reference
10
Related Paper
Citation Trend
Abstract:
Abstract Elucidating the 3D nanoscale structure of tissues and cells is essential for understanding the complexity of biological processes. Electron microscopy (EM) offers the resolution needed for reliable interpretation, but the limited throughput of electron microscopes has hindered its ability to effectively image large volumes. We report a workflow for volume EM with FAST-EM, a novel multibeam scanning transmission electron microscope that speeds up acquisition by scanning the sample in parallel with 64 electron beams. FAST-EM makes use of optical detection to separate the signals of the individual beams. The acquisition and 3D reconstruction of ultrastructural data from multiple biological samples is demonstrated. The results show that the workflow is capable of producing large reconstructed volumes with high resolution and contrast to address biological research questions within feasible acquisition time frames.Keywords:
Sample (material)
Biological specimen
Characterization
Acceleration voltage
Tomographic reconstruction
Biological specimen
Dark field microscopy
Electron scattering
Cite
Citations (0)
Richard Feynman pointed out in his 1959 speech, “It would be very easy to make an analysis of any complicated chemical substance; all one would have to do would be to look at it and see where the atoms are… I put this out as a challenge: Is there no way to make the electron microscope more powerful?” Over more than 50 years, with significant achievements in electron microscope and the developments of aberration-corrected electron optics, scanning transmission electron microscopy (STEM) has reached spatial resolution of 0.5 Å. Recently, in combination of the state-of-the-art scanning transmission electron microscope (STEM) and a tomographic reconstruction technique known as equally sloped tomography (EST), we have achieved electron tomography at 2.4 angstrom resolution [ 1 ]. This general method has been applied to reveal almost all atoms inside a Pt nanoparticle with unprecedented details [ 2 ]. Most recently, Feynman's 1959 challenge has been tackled by successfully determining 3D coordinates of 3,769 individual atoms at the tip of a tungsten needle with a precision of ~19 picometers [ 3 ]. In this talk, I will present the achievements of this novel technique.
Tomographic reconstruction
Cite
Citations (0)
Electron tomography is essential for investigating the three-dimensional (3D) structure of nanomaterials. However, many of these materials, such as metal–organic frameworks (MOFs), are extremely sensitive to electron radiation, making it difficult to acquire a series of projection images for electron tomography without inducing electron-beam damage. Another significant challenge is the high contrast in high-angle annular dark field scanning transmission electron microscopy that can be expected for nanocomposites composed of a metal nanoparticle and an MOF. This strong contrast leads to so-called metal artifacts in the 3D reconstruction. To overcome these limitations, we here present low-dose electron tomography based on four-dimensional scanning transmission electron microscopy (4D-STEM) data sets, collected using an ultrafast and highly sensitive direct electron detector. As a proof of concept, we demonstrate the applicability of the method for an Au nanostar embedded in a ZIF-8 MOF, which is of great interest for applications in various fields, including drug delivery.
Nanomaterials
Dark field microscopy
Cite
Citations (3)
The scanning-transmission imaging mode in the SEM allows for the threedimensional tomographic reconstruction of a specimen, starting from a set of projection images. Compressed sensing was used to solve the undetermined problem of structure reconstruction and was proven capable of overcoming the limitations arising from the sampling scheme. Reconstructions of cobalt particles within a carbon nanotube and collagen fibrils in a dermal tissue are presented, demonstrating the potential of this technique in the set of 3-D electron microscopy methods for both physical and biological science.
Tomographic reconstruction
Biological specimen
Cite
Citations (4)
Acceleration voltage
Tomographic reconstruction
Cryo-electron tomography
Cite
Citations (33)
Palladium nanoparticles, with potential interest for various applications in catalysis, have been studied by Transmission Electron Microscopy (TEM). In order to ascertain the exact morphology of these particles, electron tomography was perfomed in the Scanning TEM, High Angle Annular Dark Field (STEM–HAADF) imaging mode. Several geometrical forms have thus been characterized three-dimensionally at the nanometre scale, among them pentagonal rods and more complex bipyramidal nanocrystals.
Dark field microscopy
Nanometre
Rod
Cite
Citations (19)
Electron tomography offers useful three-dimensional (3D) structural information, which cannot be observed by two-dimensional imaging. By combining annular dark-field scanning transmission electron microscopy (ADF STEM) with aberration correction, the resolution of electron tomography has reached atomic resolution. However, tomography based on ADF STEM inherently suffers from several issues, including a high electron-dose requirement, poor contrast for light elements, and artifacts from image-contrast nonlinearity. Here, we develop an alternative method called multislice electron tomography (MSET) based on four-dimensional STEM tilt series. Our simulations show that multislice-based 3D reconstruction can effectively reduce undesirable reconstruction artifacts from the nonlinear contrast, allowing precise determination of atomic structures with improved sensitivity for low-Z elements, at considerably low electron-dose conditions. We expect that the MSET method can be applied to a wide variety of materials, including radiation-sensitive samples and materials containing light elements whose 3D atomic structures have never been fully elucidated due to electron-dose limitations or nonlinear imaging contrast.
Multislice
Dark field microscopy
Cite
Citations (14)
Apart from describing the occurrence and detailed crystallographic nature of novel five-fold twinned nanoparticles (<5 nm) of the selective hydrogenation catalyst GaPd2, the main thrust of this work is to demonstrate a method of characterizing, by electron tomography, the structural morphologies of large agglomerates, consisting of ca. 1800 nanoparticles, the individual sizes of which fall in the range of 1–30 nm in equivalent diameter. The so-called segmentation of electron tomograms, usually evaluated manually and vulnerable to subjective bias (as well as being laborious) is carried out by utilizing sophisticated, yet readily implementable, image processing techniques that facilitate versatile 3D nanometrological analysis. Such procedures will play an important role in the move toward quantitative 3D characterization at the nanoscale and are applicable to numerous other systems of technological and catalytic interest (such as fuel-cell electrodes) where there are agglomerates of nanoparticles and nanoclusters of various compositions, including those that are supported on high-surface-area solids.
Nanoclusters
Agglomerate
Dark field microscopy
Characterization
Photoemission electron microscopy
Cite
Citations (38)
The spatial resolution of transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs) has been drastically improved by introducing aberration correction. However, observable range in electron sensitive zeolites are still limited due to electron irradiation damages. Nevertheless, atomic resolution imaging of some zeolites is currently being realized by various developments in electron microscopic hardware, such as high sensitivity cameras. On the other hand, surveying the status of TEM and STEM imaging is very important for further progress in the structural analysis of zeolites. Here, we demonstrate and compare the high-resolution imaging of zeolites with several kinds of imaging modes.
Atomic units
Contrast transfer function
Cite
Citations (0)
Electron tomography is a well-established technique for three-dimensional structure determination of (almost) amorphous specimens in life sciences applications. With the recent advances in nanotechnology and the semiconductor industry, there is also an increasing need for high-resolution three-dimensional (3D) structural information in physical sciences. In this article, we evaluate the capabilities and limitations of transmission electron microscopy (TEM) and high-angle-annular-dark-field scanning transmission electron microscopy (HAADF-STEM) tomography for the 3D structural characterization of partially crystalline to highly crystalline materials. Our analysis of catalysts, a hydrogen storage material, and different semiconductor devices shows that features with a diameter as small as 1-2 nm can be resolved in three dimensions by electron tomography. For partially crystalline materials with small single crystalline domains, bright-field TEM tomography provides reliable 3D structural information. HAADF-STEM tomography is more versatile and can also be used for high-resolution 3D imaging of highly crystalline materials such as semiconductor devices.
Dark field microscopy
Characterization
Cite
Citations (237)