Towards an optimum design for thin film phase plates.

A variety of physical phase plate designs have been developed to maximize phase contrast for weak phase objects in the transmission electron microscope (TEM). Most progress towards application in structural biology has been made with Zernike PPs consisting of a ~30 nm film of amorphous carbon with a central hole. Although problems such as beam-induced deterioration of Zernike PPs remain unsolved, it is likely that thin film phase plates will be applied routinely in TEM of ice-embedded biological specimens in the near future. However, the thick carbon film of thin film PPs dampens high-resolution information, which precludes their use for single-particle electron cryo-microscopy at atomic resolution. In this work, an improved design for a thin film phase plate is proposed, combining the advantages of Zernike PPs and 2D materials, such as graphene. The improved design features a disc of phase-shifting material mounted on an ultrathin support film. The proposed device imparts a phase shift only to electrons scattered to low angles, whereas contrast at high resolution is generated by conventional defocusing. The device maximizes phase contrast at low spatial frequencies, where defocus contrast is limiting, while damping of information at high spatial frequencies is avoided. Experiments demonstrate that the fabrication of such a device is feasible.