X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded1-3. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction ‘snapshots’ are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source4. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes5. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (~200 nm to 2 μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes6. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.

Abstract. The European Photon Imaging Camera (EPIC) consortium has provided the focal plane instruments for the three X-ray mirror systems on XMM-Newton. Two cameras with a reflecting grating spectrometer in the optical path are equipped with MOS type CCDs as focal plane detectors (Turner 2001), the telescope with the full photon flux operates the novel pn-CCD as an imaging X-ray spectrometer. The pn-CCD camera system was developed under the leadership of the Max-Planck-Institut für extraterrestrische Physik (MPE), Garching. The concept of the pn-CCD is described as well as the different operational modes of the camera system. The electrical, mechanical and thermal design of the focal plane and camera is briefly treated. The in-orbit performance is described in terms of energy resolution, quantum efficiency, time resolution, long term stability and charged particle background. Special emphasis is given to the radiation hardening of the devices and the measured and expected degradation due to radiation damage of ionizing particles in the first 9 months of in orbit operation.Key words. XMM-Newton -back illuminated pn-CCDs -radiation hardness -energy resolution -quantum efficiency -particle and flourescence background

X-ray lasers offer new capabilities in understanding the structure of biological systems, complex materials and matter under extreme conditions1–4. Very short and extremely bright, coherent X-ray pulses can be used to outrun key damage processes and obtain a single diffraction pattern from a large macromolecule, a virus or a cell before the sample explodes and turns into plasma1. The continuous diffraction pattern of non-crystalline objects permits oversampling and direct phase retrieval2. Here we show that high-quality diffraction data can be obtained with a single X-ray pulse from a non-crystalline biological sample, a single mimivirus particle, which was injected into the pulsed beam of a hard-X-ray free-electron laser, the Linac Coherent Light Source5. Calculations indicate that the energy deposited into the virus by the pulse heated the particle to over 100,000 K after the pulse had left the sample. The reconstructed exit wavefront (image) yielded 32-nm full-period resolution in a single exposure and showed no measurable damage. The reconstruction indicates inhomogeneous arrangement of dense material inside the virion. We expect that significantly higher resolutions will be achieved in such experiments with shorter and brighter photon pulses focused to a smaller area. The resolution in such experiments can be further extended for samples available in multiple identical copies.