Development of an ultrafast pulsed ponderomotive phase plate for cryo-electron tomography

Abstract

AbstractCryo-electron tomography (cryo-ET) is a powerful modality for resolving cellular structures in their native state. While single-particle cryo-electron microscopy (cryo-EM) excels in determining protein structures purified from recombinant or endogenous sources, cryo-ET suffers from low contrast in crowded cellular milieux. A novel experimental approach to enhance contrast in cryo-ET is to manipulate the phase of scattered pulsed electrons using ultrafast pulsed photons. Here, we outline the experimental design of a proof-of-concept electron microscope and demonstrate synchronization between electron packets and laser pulses. Further, we show ultrabright photoemission of electrons from an alloy field emission tip using femtosecond ultraviolet pulses. These experiments pave the way towards exploring the utility of the ponderomotive effect using pulsed radiation to increase phase contrast in cryo-ET of subcellular protein complexesin situ, thus advancing the field of cell biology.