Sebastian Schorb

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.

X-ray–optical cross-correlator for gas-phase experiments at the Linac Coherent Light Source free-electron laser

X-ray–optical pump–probe experiments at the Linac Coherent Light Source (LCLS) have so far been limited to a time resolution of 280 fs fwhm due to timing jitter between the accelerator-based free-electron laser (FEL) and optical lasers. We have implemented a single-shot cross-correlator for femtosecond x-ray and infrared pulses. A reference experiment relying only on the pulse arrival time information from the cross-correlator shows a time resolution better than 50 fs fwhm (22 fs rms) and also yields a direct measurement of the maximal x-ray pulse length. The improved time resolution enables ultrafast pump–probe experiments with x-ray pulses from LCLS and other FEL sources.

X-Ray Diffraction from Isolated and Strongly Aligned Gas-Phase Molecules with a Free-Electron Laser

We report experimental results on x-ray diffraction of quantum-state-selected and strongly aligned ensembles of the prototypical asymmetric rotor molecule 2,5-diiodobenzonitrile using the Linac Coherent Light Source. The experiments demonstrate first steps toward a new approach to diffractive imaging of distinct structures of individual, isolated gas-phase molecules. We confirm several key ingredients of single molecule diffraction experiments: the abilities to detect and count individual scattered x-ray photons in single shot diffraction data, to deliver state-selected, e.g., structural-isomer-selected, ensembles of molecules to the x-ray interaction volume, and to strongly align the scattering molecules. Our approach, using ultrashort x-ray pulses, is suitable to study ultrafast dynamics of isolated molecules.

Ultra-fast and ultra-intense x-ray sciences: first results from the Linac Coherent Light Source free-electron laser

X-ray free-electron lasers (FELs) produce femtosecond x-ray pulses with unprecedented intensities that are uniquely suited for studying many phenomena in atomic, molecular, and optical (AMO) physics. A compilation of the current developments at the Linac Coherent Light Source (LCLS) and future plans for the LCLS-II and Next Generation Light Source (NGLS) are outlined. The AMO instrumentation at LCLS and its performance parameters are summarized. A few selected experiments representing the rapidly developing field of ultra-fast and peak intensity x-ray AMO sciences are discussed. These examples include fundamental aspects of intense x-ray interaction with atoms, nonlinear atomic physics in the x-ray regime, double core-hole spectroscopy, quantum control experiments with FELs and ultra-fast x-ray induced dynamics in clusters. These experiments illustrate the fundamental aspects of the interaction of intense short pulses of x-rays with atoms, molecules and clusters that are probed by electron and ion spectroscopies as well as ultra-fast x-ray scattering.

Shapes and vorticities of superfluid helium nanodroplets

X-raying superfluid helium droplets When physicists rotate the superfluid 4He, it develops a regular array of tiny whirlpools, called vortices. The same phenomenon should occur in helium droplets half a micrometer in size, but studying individual droplets is tricky. Gomez et al. used x-ray diffraction to deduce the shape of individual rotating droplets and image the resulting vortex patterns, which confirmed the superfluidity of the droplets. They found that superfluid droplets can host a surprising number of vortices and can rotate faster than normal droplets without disintegrating. Science, this issue p. 906 Vortex lattices inside individual helium droplets are imaged using x-ray diffraction. Helium nanodroplets are considered ideal model systems to explore quantum hydrodynamics in self-contained, isolated superfluids. However, exploring the dynamic properties of individual droplets is experimentally challenging. In this work, we used single-shot femtosecond x-ray coherent diffractive imaging to investigate the rotation of single, isolated superfluid helium-4 droplets containing ~108 to 1011 atoms. The formation of quantum vortex lattices inside the droplets is confirmed by observing characteristic Bragg patterns from xenon clusters trapped in the vortex cores. The vortex densities are up to five orders of magnitude larger than those observed in bulk liquid helium. The droplets exhibit large centrifugal deformations but retain axially symmetric shapes at angular velocities well beyond the stability range of viscous classical droplets.

Imaging charge transfer in iodomethane upon x-ray photoabsorption

Tightly tracking charge migration Electron transfer dynamics underlie many chemical and biochemical reactions. Erk et al. examined the charge migration between individual carbon and iodine atoms during dissociation of iodomethane (ICH3) molecules (see the Perspective by Pratt). After initiating scission of the C-I bond with a relatively low-energy laser pulse, they introduced a higher-energy x-ray pulse to instigate ionization and charge migration. Delaying the arrival time of the x-ray pulse effectively varied the separation distance being probed as the fragments steadily drifted apart. The experimental approach should also prove useful for future studies of charge transfer dynamics in different molecular or solid-state systems. Science, this issue p. 288; see also p. 267 A free-electron laser enables precise tracking of electron movement between segments of a dissociating molecule. [Also see Perspective by Pratt] Studies of charge transfer are often hampered by difficulties in determining the charge localization at a given time. Here, we used ultrashort x-ray free-electron laser pulses to image charge rearrangement dynamics within gas-phase iodomethane molecules during dissociation induced by a synchronized near-infrared (NIR) laser pulse. Inner-shell photoionization creates positive charge, which is initially localized on the iodine atom. We map the electron transfer between the methyl and iodine fragments as a function of their interatomic separation set by the NIR–x-ray delay. We observe signatures of electron transfer for distances up to 20 angstroms and show that a realistic estimate of its effective spatial range can be obtained from a classical over-the-barrier model. The presented technique is applicable for spatiotemporal imaging of charge transfer dynamics in a wide range of molecular systems.

Spectral encoding of x-ray/optical relative delay.

We present a new technique for measuring the relative delay between a soft x-ray FEL pulse and an optical laser that indicates a sub 25 fs RMS measurement error. An ultra-short x-ray pulse photo-ionizes a semiconductor (Si(3)N(4)) membrane and changes the optical transmission. An optical continuum pulse with a temporally chirped bandwidth spanning 630 nm-710 nm interacts with the membrane such that the timing of the x-ray pulse can be determined from the onset of the spectral modulation of the transmitted optical pulse. This experiment demonstrates a nearly in situ single-shot measurement of the x-ray pulse arrival time relative to the ultra-short optical pulse.

The 3D-architecture of individual free silver nanoparticles captured by X-ray scattering

The diversity of nanoparticle shapes generated by condensation from gaseous matter reflects the fundamental competition between thermodynamic equilibration and the persistence of metastable configurations during growth. In the kinetically limited regime, intermediate geometries that are favoured only in early formation stages can be imprinted in the finally observed ensemble of differently structured specimens. Here we demonstrate that single-shot wide-angle scattering of femtosecond soft X-ray free-electron laser pulses allows three-dimensional characterization of the resulting metastable nanoparticle structures. For individual free silver particles, which can be considered frozen in space for the duration of photon exposure, both shape and orientation are uncovered from measured scattering images. We identify regular shapes, including species with fivefold symmetry and surprisingly large aspect ratio up to particle radii of the order of 100 nm. Our approach includes scattering effects beyond Born’s approximation and is remarkably efficient—opening up new routes in ultrafast nanophysics and free-electron laser science.

Femtosecond x-ray photoelectron diffraction on gas-phase dibromobenzene molecules

We present time-resolved femtosecond photoelectron momentum images and angular distributions of dissociating, laser-aligned 1,4-dibromobenzene (C6H4Br2) molecules measured in a near-infrared pump, soft-x-ray probe experiment performed at an x-ray free-electron laser. The observed alignment dependence of the bromine 2p photoelectron angular distributions is compared to density functional theory calculations and interpreted in terms of photoelectron diffraction. While no clear time-dependent effects are observed in the angular distribution of the Br(2p) photoelectrons, other, low-energy electrons show a pronounced dependence on the time delay between the near-infrared laser and the x-ray pulse.