A Method for High-Resolution Three-Dimensional Reconstruction with Ewald Sphere Curvature Correction from Transmission Electron Images. (October 2022)

• Record Type: Journal Article
• Authors: Gureyev, Timur E.; Paganin, David M.; Brown, Hamish G.; Quiney, Harry M.; Allen, Leslie J.
• Is Part Of: Microscopy and microanalysis. Volume 28:Number 5(2022)
• Page Start: 1550
• Page End: 1566
• Publication Date: 2022-10
• Subjects: atomic resolution -- cryo-EM -- Ewald sphere curvature -- single-particle analysis -- transmission electron microscopy; Microscopy -- Periodicals; Microchemistry -- Periodicals; 502.82
• DOI: 10.1017/S1431927622000630
• Languages: English
• ISSNs: 1431-9276

Abstract

Abstract: Abstract : A method for three-dimensional reconstruction of objects from defocused images collected at multiple illumination directions in high-resolution transmission electron microscopy is presented. The method effectively corrects for the Ewald sphere curvature by taking into account the in-particle propagation of the electron beam. Numerical simulations demonstrate that the proposed method is capable of accurately reconstructing biological molecules or nanoparticles from high-resolution defocused images under conditions achievable in single-particle electron cryo-microscopy or electron tomography with realistic radiation doses, non-trivial aberrations, multiple scattering, and other experimentally relevant factors. The physics of the method is based on the well-known Diffraction Tomography formalism, but with the phase-retrieval step modified to include a conjugation of the phase (i.e., multiplication of the phase by a negative constant). At each illumination direction, numerically backpropagating the beam with the conjugated phase produces maximum contrast at the location of individual atoms in the molecule or nanoparticle. The resultant algorithm, Conjugated Holographic Reconstruction, can potentially be incorporated into established software tools for single-particle analysis, such as, for example, RELION or FREALIGN, in place of the conventional contrast transfer function correction procedure, in order to account for the Ewald sphere curvature and improve the spatial resolution of the three-dimensional reconstruction.