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Published on: July 29, 2021
Paraxial charge compensator for electron cryomicroscopy
John A Berriman1, Peter B Rosenthal
1Division of Physical Biochemistry, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London, NW7 1AA, United Kingdom.
Abstract:
We describe a multi-hole condenser aperture for the production of several electron beams in the transmission electron microscope (TEM) making it possible to simultaneously image and irradiate spatially separated regions of a specimen. When the specimen is a thin film of vitreous ice suspended over a holey carbon film, simultaneous irradiation of the adjacent carbon support with the off-axis beam compensates for some of the effects of charging in the image formed by a beam irradiating only the ice. Because the intervening region is not irradiated, charge-neutralization of frozen-hydrated specimens can occur by a through-space mechanism such as the emission of secondary electrons from a grounded carbon support film. We use paraxial charge compensation (PCC) to control the amount of charge build-up on the specimen and observe the effects of charge on images. The multi-hole aperture thus provides a tool for investigating the mechanism of charging and charge mitigation during the imaging of radiation sensitive biological specimens by cryomicroscopy.
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