Radiation dose reduction and image enhancement in biological imaging through equally-sloped tomography
Year: 2008
Type: article
Abstract: Electron tomography is currently the highest resolution imaging modality available to study the 3D structures of pleomorphic macromolecular assemblies, viruses, organelles and cells. Unfortunately, the resolution is currently limited to 3-5nm by several factors including the dose tolerance of biological specimens and the inaccessibility of certain tilt angles. Here we report the first experimental demonstration of equally-sloped tomography (EST) to alleviate these problems. As a proof of principle, we applied EST to reconstructing frozen-hydrated keyhole limpet hemocyanin molecules from a tilt-series taken with constant slope increments. In comparison with weighted back-projection (WBP), the algebraic reconstruction technique (ART) and the simultaneous algebraic reconstruction technique (SART), EST reconstructions exhibited higher contrast, less peripheral noise, more easily detectable molecular boundaries and reduced missing wedge effects. More importantly, EST reconstructions including only two-thirds the original images appeared to have the same resolution as full WBP reconstructions, suggesting that EST can either reduce the dose required to reach a given resolution or allow higher resolutions to be achieved with a given dose. EST was also applied to reconstructing a frozen-hydrated bacterial cell from a tilt-series taken with constant angular increments. The results confirmed similar benefits when standard tilts are utilized.
Source: Journal of Structural Biology
Language: English
Subfield: Structural Biology
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,350
Funders: National Science Foundation, U.S. Department of Energy, Alfred P. Sloan Foundation, Gordon and Betty Moore Foundation, California Institute of Technology, Agouron Institute, National Institutes of Health, Beckman Institute, California Institute of Technology, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, Division of Materials Research, Basic Energy Sciences
Awards: G201454641, MRI: Development Of A Next-Generation Coherent X-Ray Diffraction Microscope For 3d Imaging Of Nanosclae Systems, G2043586680, G222768804, G2769404516, G3284393805, Center for the Structural Biology of Cellular Host Elements in Egress, Trafficking, and Assembly of HIV (CHEETAH Center), G3906547125, PROBING SINGLE MICROBIAL PROTEINS AND MULTI-PROTEIN COMPLEXES WITH BIOCONJUGATED QUANTUM DOTS, G4515935217, G5462169360, G5648016710, Three-Dimensional Imaging of Whole Cells by Using X-ray Diffraction Microscopy, Structure and function of the prokaryotic cytoskeleton by electron cryotomography, G6605472885, THREE DIMENSIONAL IMAGING OF NANOSCALE MATERIALS BY USING COHERENT X-RAYS, G7967587074, G8177302771, G882236735, G8985739490, G9000040526
OpenAlex ID: W2165187677