Bragg coherent diffraction imaging and metrics for radiation damage in protein micro-crystallography.

The proliferation of extremely intense synchrotron sources has enabled ever higher-resolution structures to be obtained using data collected from smaller and often more imperfect biological crystals (Helliwell, 1984). Synchrotron beamlines now exist that are capable of measuring data from single crystals that are just a few micrometres in size. This provides renewed motivation to study and understand the radiation damage behaviour of small protein crystals. Reciprocal-space mapping and Bragg coherent diffractive imaging experiments have been performed on cryo-cooled microcrystals of hen egg-white lysozyme as they undergo radiation damage. Several well established metrics, such as intensity-loss and lattice expansion, are applied to the diffraction data and the results are compared with several new metrics that can be extracted from the coherent imaging experiments. Individually some of these metrics are inconclusive. However, combining metrics, the results suggest that radiation damage behaviour in protein micro-crystals differs from that of larger protein crystals and may allow them to continue to diffract for longer. A possible mechanism to account for these observations is proposed.

Expected resolution limits of x-ray free-electron laser single-particle imaging for realistic source and detector properties.

The unprecedented intensity of x-ray free-electron laser sources has enabled single-particle x-ray diffraction imaging (SPI) of various biological specimens in both two-dimensional projection and three dimensions (3D). The potential of studying protein dynamics in their native conditions, without crystallization or chemical staining, has encouraged researchers to aim for increasingly higher resolutions with this technique. The currently achievable resolution of SPI is limited to the sub-10 nanometer range, mainly due to background effects, such as instrumental noise and parasitic scattering from the carrier gas used for sample delivery. Recent theoretical studies have quantified the effects of x-ray pulse parameters, as well as the required number of diffraction patterns to achieve a certain resolution, in a 3D reconstruction, although the effects of detector noise and the random particle orientation in each diffraction snapshot were not taken into account. In this work, we show these shortcomings and address limitations on achievable image resolution imposed by the adaptive gain integrating pixel detector noise.

Energy-resolved neutron imaging options at a small angle neutron scattering instrument at the Australian Center for Neutron Scattering.

Energy-resolved neutron imaging experiments conducted on the Small Angle Neutron Scattering (SANS) instrument, Bilby, demonstrate how the capabilities of this instrument can be enhanced by a relatively simple addition of a compact neutron counting detector. Together with possible SANS sample surveying and location of the region of interest, this instrument is attractive for many imaging applications. In particular, the combination of the cold spectrum of the neutron beam and its pulsed nature enables unique non-destructive studies of the internal structure for samples that are opaque to other more traditional techniques. In addition to conventional white beam neutron radiography, we conducted energy-resolved imaging experiments capable of resolving features related to microstructure in crystalline materials with a spatial resolution down to ∼0.1 mm. The optimized settings for the beamline configuration were determined for the imaging modality, where the compromise between the beam intensity and the achievable spatial resolution is of key concern.