Abstract
Cryo-electron microscopy is an essential tool for high-resolution structural studies of biological systems. This method relies on the use of phase contrast imaging at high defocus to improve information transfer at low spatial frequencies at the expense of higher spatial frequencies. Here we demonstrate that electron ptychography can recover the phase of the specimen with continuous information transfer across a wide range of the spatial frequency spectrum, with improved transfer at lower spatial frequencies, and as such is more efficient for phase recovery than conventional phase contrast imaging. We further show that the method can be used to study frozen-hydrated specimens of rotavirus double-layered particles and HIV-1 virus-like particles under low-dose conditions (5.7 e/Å2) and heterogeneous objects in an Adenovirus-infected cell over large fields of view (1.14 × 1.14 μm), thus making it suitable for studies of many biologically important structures.
Highlights
Cryo-electron microscopy is an essential tool for high-resolution structural studies of biological systems
Electron ptychographic data was recorded in a scanning electron diffraction mode as shown schematically in Fig. 1a at 300 kV using a double aberration corrected JEOL ARM300CF
Rotavirus double-layered particles (DLPs) with a diameter of 76.5 nm and immature HIV-1 virus-like particles (VLPs) with variable sizes were used to demonstrate this method under cryo-conditions[38]
Summary
Cryo-electron microscopy is an essential tool for high-resolution structural studies of biological systems. In order to compare information transfer using ptychography and conventional phase contrast imaging at high defocus, power spectra and corresponding radial averages were calculated (Fig. 1d, g).
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