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Characterizing intense, focused x-ray free electron laser (FEL) pulses is crucial for their use in diffractive imaging. We describe how the distribution of average phase tilts and intensities on hard x-ray pulses with peak intensities of 10(21) W/m(2) can be retrieved from an ensemble of diffraction patterns produced by 70 nm-radius polystyrene spheres, in a manner that mimics wavefront sensors. Besides showing that an adaptive geometric correction may be necessary for diffraction data from randomly injected sample sources, our paper demonstrates the possibility of collecting statistics on structured pulses using only the diffraction patterns they generate and highlights the imperative to study its impact on single-particle diffractive imaging.

Free-electron lasers (FEL) hold the potential to revolutionize structural biology by producing X-ray pules short enough to outrun radiation damage, thus allowing imaging of biological samples without the limitation from radiation damage. Thus, a major part of the scientific case for the first FELs was three-dimensional (3D) reconstruction of non-crystalline biological objects. In a recent publication we demonstrated the first 3D reconstruction of a biological object from an X-ray FEL using this technique. The sample was the giant Mimivirus, which is one of the largest known viruses with a diameter of 450 nm. Here we present the dataset used for this successful reconstruction. Data-analysis methods for single-particle imaging at FELs are undergoing heavy development but data collection relies on very limited time available through a highly competitive proposal process. This dataset provides experimental data to the entire community and could boost algorithm development and provide a benchmark dataset for new algorithms.

We demonstrate the use of an X-ray free electron laser synchronized with an optical pump laser to obtain X-ray diffraction snapshots from the photoactivated states of large membrane protein complexes in the form of nanocrystals flowing in a liquid jet. Light-induced changes of Photosystem I-Ferredoxin co-crystals were observed at time delays of 5 to 10 µs after excitation. The result correlates with the microsecond kinetics of electron transfer from Photosystem I to ferredoxin. The undocking process that follows the electron transfer leads to large rearrangements in the crystals that will terminally lead to the disintegration of the crystals. We describe the experimental setup and obtain the first time-resolved femtosecond serial X-ray crystallography results from an irreversible photo-chemical reaction at the Linac Coherent Light Source. This technique opens the door to time-resolved structural studies of reaction dynamics in biological systems.

Abstract: In the MPI-Halbleiterlabor (MPI-HLL) we are developing fully depleted pn-type Charge Coupled Devices (pnCCD) with a thick radiation-sensitive region. Since 1997, the mesh experiment has been established as an unique technique to determine an X-ray interaction position with subpixel resolution. In order to investigate the signal charge behavior in pnCCDs, we performed the mesh experiment using various characteristic X-ray lines. Cu-L (0.9keV), Al-K (1.5keV) and Ti-K (4.5keV) were used and the device operated under various conditions, e.g. register voltages. We obtained the restored pixel images for various X-ray event patterns. The photon entrance window of the back illuminated pnCCD shows highly uniform detection efficiency within a pixel. We measured the charge cloud size generated by an X-ray photon inside the CCD. The measured charge cloud sizes are around 6 m (rms) depending on the X-ray energy. The operation with different register conditions revealed asymmetries and regions with a weaker electric field around pixel boundaries. [Copyright &y& Elsevier]