Thick (3D) Sample Imaging Using iDPC-STEM at Atomic Scale

Lazić, Ivan; Bosch, Eric; Yücelen, Emrah; Imlau, Robert; Lazar, Sorin

doi:10.1017/s1431927618001344

Cited by 7 publications

(9 citation statements)

References 7 publications

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“…When increasing the CSA and the probe is aberration-free, higher resolution is obtained while the beam depth of focus becomes smaller. Consequently, when the sample thickness exceeds the decreasing beam-waist length, the images represent optical depth sections rather than projections 50 , 53 . We estimate that the sample regions imaged here have thicknesses of 18 to 54 nm corresponding to a diameter of a single TMV rod and three TMV rods, respectively.…”

Section: Discussionmentioning

confidence: 99%

Single-particle cryo-EM structures from iDPC–STEM at near-atomic resolution

et al. 2022

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In electron cryomicroscopy (cryo-EM), molecular images of vitrified biological samples are obtained by conventional transmission microscopy (CTEM) using large underfocuses and subsequently computationally combined into a high-resolution three-dimensional structure. Here, we apply scanning transmission electron microscopy (STEM) using the integrated differential phase contrast mode also known as iDPC–STEM to two cryo-EM test specimens, keyhole limpet hemocyanin (KLH) and tobacco mosaic virus (TMV). The micrographs show complete contrast transfer to high resolution and enable the cryo-EM structure determination for KLH at 6.5 Å resolution, as well as for TMV at 3.5 Å resolution using single-particle reconstruction methods, which share identical features with maps obtained by CTEM of a previously acquired same-sized TMV data set. These data show that STEM imaging in general, and in particular the iDPC–STEM approach, can be applied to vitrified single-particle specimens to determine near-atomic resolution cryo-EM structures of biological macromolecules.

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Section: Discussionmentioning

confidence: 99%

Single-particle cryo-EM structures from iDPC–STEM at near-atomic resolution

et al. 2022

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“…When increasing the CSA, higher resolution is obtained provided the probe is aberration free while the beam depth of focus becomes smaller. Consequently, when sample thickness exceeds the decreasing beam-waist length, the images represent optical depth sections rather than projections [45,48]. We estimate that the sample regions imaged here have thicknesses of 18 to 54 nm corresponding to a diameter of a single TMV rod and three TMV rods, respectively.…”

Section: Discussionmentioning

confidence: 95%

Single-particle cryo-EM structures from iDPC-STEM at near-atomic resolution

Lazić

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Leidl

et al. 2021

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Electron cryo-microscopy (cryo-EM) is becoming one of the routine tools for structure determination of biological macromolecules. Commonly, molecular images are obtained by conventional transmission electron microcopy (CTEM) using underfocus and subsequently computationally combined into a high-resolution 3D structure. Here, we apply scanning transmission electron microscopy (STEM) using the integrated differential phase contrast mode also known as iDPC-STEM to the cryo-EM test specimen of tobacco mosaic virus (TMV). The micrographs show complete contrast transfer to high resolution and enable the cryo-EM structure determination at 3.5 Angstrom resolution using single-particle based helical reconstruction. A series of cryo-EM TMV maps was resolved at near-atomic resolution taken at different convergence semi-angle (CSA) beams and share identical features with maps obtained by CTEM of a previously acquired same-sized TMV data set. The associated map B-factors from iDPC-STEM match those obtained by CTEM recordings using 2nd generation direct electron detection devices. These data show that STEM imaging in general, and in particular the iDPC-STEM approach, can be applied to vitrified single-particle specimens to determine near-atomic resolution cryo-EM structures of biological macromolecules.

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“…The value of the defocus is only limited by the specimen thickness, as it is the depth at which the projected electrostatic potential is probed by the converged beam waist allowing atomically resolved 3D iDPC imaging (49). In addition, it shows enhanced contrast between the hydrogen and titanium columns as plotted in Figure S11C.…”

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confidence: 99%

“…List of the electron detectors and their corresponding collection angles that are used in the experiment and simulation. References (42)(43)(44)(45)(46)(47)(48)(49)(50)(51)…”

Section: Supplementary Materialsmentioning

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Resolving hydrogen atoms at metal-metal hydride interfaces

et al. 2020

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Hydrogen as a fuel can be stored safely with high volumetric density in metals. It can, however, also be detrimental to metals causing embrittlement. Understanding fundamental behavior of hydrogen at atomic scale is key to improve the properties of metalmetal hydride systems. However, currently, there is no robust technique capable of visualizing hydrogen atoms. Here, we demonstrate that hydrogen atoms can be imaged unprecedentedly with integrated differential phase contrast, a recently developed technique performed in a scanning transmission electron microscope. Images of the titanium-titanium monohydride interface reveal remarkable stability of the hydride phase, originating from the interplay between compressive stress and interfacial coherence. We also uncovered, thirty years after three models were proposed, which one describes the position of the hydrogen atoms with respect to the interface. Our work enables novel research on hydrides and is extendable to all materials containing light and heavy elements, including oxides, nitrides, carbides and borides.

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Thick (3D) Sample Imaging Using iDPC-STEM at Atomic Scale

Cited by 7 publications

References 7 publications

Single-particle cryo-EM structures from iDPC–STEM at near-atomic resolution

Single-particle cryo-EM structures from iDPC–STEM at near-atomic resolution

Single-particle cryo-EM structures from iDPC-STEM at near-atomic resolution

Resolving hydrogen atoms at metal-metal hydride interfaces

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