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Since the first successful serial crystallography (SX) experiment at a synchrotron radiation source, the popularity of this approach has continued to grow showing that third-generation synchrotrons can be viable alternatives to scarce X-ray free-electron laser sources. Synchrotron radiation flux may be increased ∼100 times by a moderate increase in the bandwidth (`pink beam' conditions) at some cost to data analysis complexity. Here, we report the first high-viscosity injector-based pink-beam SX experiments. The structures of proteinase K (PK) and A_2Aadenosine receptor (A_2AAR) were determined to resolutions of 1.8 and 4.2 Å using 4 and 24 consecutive 100 ps X-ray pulse exposures, respectively. Strong PK data were processed using existing Laue approaches, while weaker A_2AAR data required an alternative data-processing strategy. This demonstration of the feasibility presents new opportunities for time-resolved experiments with microcrystals to study structural changes in real time at pink-beam synchrotron beamlines worldwide. 

We examine the linear stability of a homogeneous gas–solid suspension of small Stokes number particles, with a moderate mass loading, subject to a simple shear flow. The modulation of the gravitational force exerted on the suspension, due to preferential concentration of particles in regions of low vorticity, in response to an imposed velocity perturbation, can lead to an algebraic instability. Since the fastest growing modes have wavelengths small compared with the characteristic length scale ( $$U_{g}/{\it\Gamma}$$ ) and oscillate with frequencies large compared with $${\it\Gamma}$$ , $$U_{g}$$ being the settling velocity and $${\it\Gamma}$$ the shear rate, we apply the WKB method, a multiple scale technique. This analysis reveals the existence of a number density mode which travels due to the settling of the particles and a momentum mode which travels due to the cross-streamline momentum transport caused by settling. These modes are coupled at a turning point which occurs when the wavevector is nearly horizontal and the most amplified perturbations are those in which a momentum wave upstream of the turning point creates a downstream number density wave. The particle number density perturbations reach a finite, but large amplitude that persists after the wave becomes aligned with the velocity gradient. The growth of the amplitude of particle concentration and fluid velocity disturbances is characterised as a function of the wavenumber and Reynolds number ( $$\mathit{Re}=U_{g}^{2}/{\it\Gamma}{\it\nu}$$ ) using both asymptotic theory and a numerical solution of the linearised equations. 

Crystal structure determination of biological macromolecules using the novel technique of serial femtosecond crystallography (SFX) is severely limited by the scarcity of X-ray free-electron laser (XFEL) sources. However, recent and future upgrades render microfocus beamlines at synchrotron-radiation sources suitable for room-temperature serial crystallography data collection also. Owing to the longer exposure times that are needed at synchrotrons, serial data collection is termed serial millisecond crystallography (SMX). As a result, the number of SMX experiments is growing rapidly, with a dozen experiments reported so far. Here, the first high-viscosity injector-based SMX experiments carried out at a US synchrotron source, the Advanced Photon Source (APS), are reported. Microcrystals (5–20 µm) of a wide variety of proteins, including lysozyme, thaumatin, phycocyanin, the human A 2A adenosine receptor (A 2A AR), the soluble fragment of the membrane lipoprotein Flpp3 and proteinase K, were screened. Crystals suspended in lipidic cubic phase (LCP) or a high-molecular-weight poly(ethylene oxide) (PEO; molecular weight 8 000 000) were delivered to the beam using a high-viscosity injector. In-house data-reduction (hit-finding) software developed at APS as well as the SFX data-reduction and analysis software suites Cheetah and CrystFEL enabled efficient on-site SMX data monitoring, reduction and processing. Complete data sets were collected for A 2A AR, phycocyanin, Flpp3, proteinase K and lysozyme, and the structures of A 2A AR, phycocyanin, proteinase K and lysozyme were determined at 3.2, 3.1, 2.65 and 2.05 Å resolution, respectively. The data demonstrate the feasibility of serial millisecond crystallography from 5–20 µm crystals using a high-viscosity injector at APS. The resolution of the crystal structures obtained in this study was dictated by the current flux density and crystal size, but upcoming developments in beamline optics and the planned APS-U upgrade will increase the intensity by two orders of magnitude. These developments will enable structure determination from smaller and/or weakly diffracting microcrystals.