Improvements in electron diffraction of frozen hydrated crystals by energy filtering and large-area single-electron detection

Abstract

Diffraction patterns of 3D protein crystals embedded in vitrious ice are critical to record. Inelastically scattered electrons almost completely superimpose the diffraction pattern of crystals if the thickness of the crystal is higher than the mean free path of electrons in the specimen. Figure 1 shows such an example of an unfiltered electron diffraction pattern from a frozen hydrated 3D catalase crystal. However, for thin 2D crystals electron diffraction has been the state of the art method to determine the Fourier amplitudes for reconstructions to atomic level, and in one case the possibility of obtaining Fourier phases from diffraction patterns has been studied. One of the main problems could be the background in the diffraction pattern due to inelastic scattering and the recording characteristics for electrons of conventional negative material.It was pointed out before, that the use of an energy filtered TEM (EFTEM) and of the Image Plate as a large area electron detector gives considerable improvement for detection of diffraction patterns.