O Jonsson

Femtosecond x-ray protein nanocrystallography

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 recorded. 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 Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. 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 processes. 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.

Single mimivirus particles intercepted and imaged with an X-ray laser

X-ray lasers offer new capabilities in understanding the structure of biological systems, complex materials and matter under extreme conditions. 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 plasma. The continuous diffraction pattern of non-crystalline objects permits oversampling and direct phase retrieval. 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 Source. 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.

Femtosecond free-electron laser x-ray diffraction data sets for algorithm development.

We describe femtosecond X-ray diffraction data sets of viruses and nanoparticles collected at the Linac Coherent Light Source. The data establish the first large benchmark data sets for coherent diffraction methods freely available to the public, to bolster the development of algorithms that are essential for developing this novel approach as a useful imaging technique. Applications are 2D reconstructions, orientation classification and finally 3D imaging by assembling 2D patterns into a 3D diffraction volume.

The New Uppsala Neutron Beam Facility

A new quasi‐monoenergetic neutron beam facility has been constructed at the The Svedberg Laboratory (TSL) in Uppsala, Sweden. Key features include an energy range of 20 to 175 MeV, high fluxes, and the possibility of large‐area fields. Besides cross‐section measurements, the new facility has been designed specifically to provide optimal conditions for testing of single‐event effects in electronics and for dosimetry development. First results of the beam characterization measurements performed in early 2004 are reported.

Single particle imaging with soft X-rays at the linac coherent light source

Results of coherent diffractive imaging experiments performed with soft X-rays (1-2 keV) at the Linac Coherent Light Source are presented. Both organic and inorganic nano-sized objects were injected into the XFEL beam as an aerosol focused with an aerodynamic lens. The high intensity and femtosecond duration of X-ray pulses produced by the Linac Coherent Light Source allow structural information to be recorded by X-ray diffraction before the particle is destroyed. Images were formed by using iterative methods to phase single shot diffraction patterns. Strategies for improving the reconstruction methods have been developed. This technique opens up exciting opportunities for biological imaging, allowing structure determination without freezing, staining or crystallization.

Mix-and-diffuse serial synchrotron crystallography

The structure of chitotriose bound to lysozyme after mixing times of 2 and 50 s was determined using a polyimide tape-drive device for mix-and-diffuse serial crystallography at a synchrotron light source.

Elastic neutron scattering at 96 MeV from {sup 12}C and {sup 208}Pb

A facility for detection of scattered neutrons in the energy interval 50-130 MeV, SCANDAL, has recently been installed at the 20-180 MeV neutron beam line of the The Svedberg Laboratory, Uppsala. Elastic neutron scattering from {sup 12}C and {sup 208}Pb has been studied at 96 MeV in the 10 deg. -70 deg. interval. The achieved energy resolution, 3.7 MeV, is about an order of magnitude better than for any previous experiment above 65 MeV incident energy. The present experiment represents the highest neutron energy where the ground state has been resolved from the first excited state in neutron scattering. A novel method for normalization of the absolute scale of the cross section has been used. The estimated normalization uncertainty, 3%, is unprecedented for a neutron-induced differential cross section measurement on a nuclear target. The results are compared with modern optical model predictions based on phenomenology or microscopic nuclear theory.

Single-shot diffraction data from the Mimivirus particle using an X-ray free-electron laser.

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.

Elastic neutron scattering at 96 MeV

A facility for detection of scattered neutrons in the energy interval 50–130 MeV, SCANDAL (SCAttered Nucleon Detection AssembLy), has recently been installed at the 20 – 180‐MeV neutron beam line of The Svedberg Laboratory, Uppsala. Elastic neutron scattering from 12C, 16O, 56Fe, 89Y, and 208Pb has been studied at 96 MeV in the 10 – 70° interval. The results from 12C and 208Pb have recently been published,6 while the data from 16O, 56Fe, and 89Y are under analysis. The achieved energy resolution, 3.7 MeV, is about an order of magnitude better than for any previous experiment above 65 MeV incident energy. The present experiment represents the highest neutron energy where the ground state has been resolved from the first excited state in neutron scattering. A novel method for normalization of the absolute scale of the cross section has been used. The estimated normalization uncertainty, 3%, is unprecedented for a neutron‐induced differential cross section measurement on a nuclear target. The results are com...

Investigation of Three‐Body Force Effects in Neutron‐Deuteron Scattering at 95 MeV

We have measured the neutron‐deuteron (nd) elastic‐scattering differential cross section at 95 MeV incident neutron energy, using both the Medley and the SCANDAL setups at TSL in Uppsala. The full angular distribution was covered by detecting recoil deuterons from thin CD2 targets, and the result was normalized to the neutron‐proton (np) cross section. Recent theories predict that three‐nucleon (3N) force effects, if present, would affect the cross section in the minimum region by about 30%. The results are compared with theoretical calculations and are well described if 3N forces are included.