Electron Tomography: Three-Dimensional Imaging with the Transmission Electron Microscope

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Ebook Electron Tomography Three Dimensional Imaging With The Transmission Electron Microscope

Luthi, D. Studer, J. Al-Amoudi, D. Dubochet, J. Herman, Fundamentals of computerised tomography, in: Image Reconstruction From Projection, second ed. Denk, H. Hashimoto, X. Zhou, C. Luo, K. Kawano, G. Thompson, A. Hughes, P. Skeldon, P. Withers, T. Marrow, A.

TomoJ: tomography software for three-dimensional reconstruction in transmission electron microscopy

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To evaluate depth resolution experimentally, the region of the CNT bundle showing lattice fringes with a spacing of 0. According to Eq. Evaluation of depth resolution. Lattice fringes, with a spacing of 0. The simulations show that there is a sample thickness limit even for light element materials, such as that used in the present experiment where the invMS can fully accommodate multiple scattering but where the WDD approach cannot.

In contrast to the work of Yang et al. This process removes both microstructural defects as well as the catalyst particles within CNTs and enhances their graphitic structure The optical sectioning results are shown in Supplementary Fig. The WDD reconstruction shows contrast variations along the depth direction and a noticeable contrast reversal in the reconstructed phase data Supplementary Fig. Hence, the contrast reversal observed in WDD optical sectioning is likely due to dynamical scattering. However, this is not the case for reconstructions using the invMS approach.

It can be seen that invMS reconstruction recovers more faithful phase data Supplementary Fig. Therefore, while for 2D imaging the WDD has produced impressive results at atomic resolution, its 3D optical sectioning capability for thick samples is limited by multiple scattering. This is consistent with the theory underlying the WDD, which makes use of the multiplicative approximation.

Overall, the invMS approach provides depth-resolved sectioning, which is robust to multiple scattering, although the attainable resolution has yet to be shown to be competitive with that achieved using the WDD and a focused probe. Compared to experiments using a focused probe and an integrating high-speed pixelated detector 30 as used to record data for the recent WDD work 19 , 29 , the defocused probe used in invMS is incompatible with the optical conditions required for annular dark-field imaging.

However, despite this disadvantage, the data acquisition described here does not necessarily require a high-speed pixelated detector 30 as used successfully in the WDD experiments 29 , the speed of which can practically limit the number of probe positions and hence the size of the final reconstruction in pixels. In future, we anticipate extending this approach by using direct counting electron detectors that are designed to have a very wide dynamic range Using this type of detector, we expect that much weaker signals can be detected from weakly scattering objects under low-dose conditions and also that the signal in the dark-field region outside the BF disk will be recorded with useable signal to noise ratio.

In this geometry, the scattering angle used in the reconstruction will be extended to the maximum collection angle acquired by the detector, which will improve both lateral and depth resolution. In conclusion, we have demonstrated for the first time that defocused probe electron ptychography using an invMS method can provide depth-sectioned information from a 3D complex transmission function of a thick sample including both amplitude and phase.

The method provides high contrast, quantitative phase maps at close to atomic lateral resolution and with a few tens of nanometers depth resolution.

Transmission Electron Tomography: Intracellular Insight for the Future of Medicine

This 3D electron ptychographic method is potentially applicable to in situ TEM experiments as it does not require mechanical tilting of the specimen through large angles as is the case for electron tomography. The additional capability to carry out post-acquisition focusing of the reconstruction coupled with high-dose efficiency 61 particularly when coupled to high-speed and high-sensitivity detectors 55 should provide a route to structural determination, using the recovered, fully quantitative ptychographic phase from thicker, radiation sensitive specimens. In the future, we anticipate that this method will find a wide range of applications in 3D structure determination of thick objects, ranging from inorganic nanostructures, heterostructures or ferroic domain structures to biological macromolecules.

This high-temperature process efficiently graphitized the multiwall CNTs In order to disperse the tubes with different geometries, dropping and drying was repeated several times. The iterative method for reconstruction used the ptychography algorithm with the invMS approach 22 is summarized below.

The multislice method is widely used in electron microscope simulations. The corresponding diffraction pattern acquired in the experiment is labeled, I j. For brevity, we subsequently omit the coordinate, r. For the next iteration, the probe is moved to the next position in the data set and the newly updated object slice is used as initial estimate.

For the experimental set-up from which data was collected, the slice thickness can be chosen such that the minimum separation makes two neighboring slices of the object lie outside of the bounds of the multiplicative approximation For a finer slice thickness and a larger total slice number, a greater number of unknown pixels in the specimen reconstruction need to be reconstructed. Therefore, both of these values are also dependent on the degree of the redundancy of the ptychographic data, similar to the over-sampling ratio as described elsewhere This experimental configuration ensures that the extent of the illumination was well defined and gives a probe diameter of about 5.

In this configuration, the overlap between adjacent positions was calculated to be From the over-sampling ratio 62 , the degree of the redundancy of ptychographic data used here is estimated as:.

Maire, E. Kisi, E. Williams, D. Spence, J. Press, Frank, J. Lucic, V. Structural studies by electron tomography: from cells to molecules. Midgley, P.

Background

Electron tomography and holography in materials science. Miao, J. Atomic electron tomography: 3D structures without crystals. Science , aaf Van Aert, S. Three-dimensional atomic imaging of crystalline nanoparticles. Nature , — Bals, S. A new approach for electron tomography: annular dark field transmission electron microscopy. Goris, B.

What is Cryo-Electron Microscopy (Cryo-EM)?