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Imaging biological macromolecules in thick specimens: The role of inelastic scattering in cryoEM.


ABSTRACT: We investigate potential improvements in using electron cryomicroscopy to image thick specimens with high-resolution phase contrast imaging. In particular, using model experiments, electron scattering theory, Monte Carlo and multislice simulations, we determine the potential for improving electron cryomicrographs of proteins within a cell using chromatic aberration (Cc) correction. We show that inelastically scattered electrons lose a quantifiable amount of spatial coherence as they transit the specimen, yet can be used to enhance the signal from thick biological specimens (in the 1000 to 5000 Å range) provided they are imaged close to focus with an achromatic lens. This loss of information quantified here, which we call "specimen induced decoherence", is a fundamental limit on imaging biological molecules in situ. We further show that with foreseeable advances in transmission electron microscope technology, it should be possible to directly locate and uniquely identify sub-100 kDa proteins without the need for labels, in a vitrified specimen taken from a cell.

SUBMITTER: Dickerson JL 

PROVIDER: S-EPMC9355893 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Imaging biological macromolecules in thick specimens: The role of inelastic scattering in cryoEM.

Dickerson Joshua L JL   Lu Peng-Han PH   Hristov Dilyan D   Dunin-Borkowski Rafal E RE   Russo Christopher J CJ  

Ultramicroscopy 20220319


We investigate potential improvements in using electron cryomicroscopy to image thick specimens with high-resolution phase contrast imaging. In particular, using model experiments, electron scattering theory, Monte Carlo and multislice simulations, we determine the potential for improving electron cryomicrographs of proteins within a cell using chromatic aberration (C<sub>c</sub>) correction. We show that inelastically scattered electrons lose a quantifiable amount of spatial coherence as they t  ...[more]

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