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Fixed-target pump–probe SFX: eliminating the scourge of light contamination

X-ray free-electron laser (XFEL) light sources have enabled the rapid growth of time-resolved structural experiments, which provide crucial information on the function of macromolecules and their mechanisms. Here, the aim was to commission the SwissMX fixed-target sample-delivery system at the SwissFEL Cristallina experimental station using the PSI-developed micro-structured polymer (MISP) chip for pump–probe time-resolved experiments. To characterize the system, crystals of the light-sensitive protein light–oxygen–voltage domain 1 (LOV1) from Chlamydomonas reinhardtii were used. Using different experimental settings, the accidental illumination, referred to as light contamination, of crystals mounted in wells adjacent to those illuminated by the pump laser was examined. It was crucial to control the light scattering from and through the solid supports otherwise significant contamination occurred. However, the results here show that the opaque MISP chips are suitable for defined pump–probe studies of a light-sensitive protein. The experiment also probed the sub-millisecond structural dynamics of LOV1 and indicated that at Δt = 10 µs a covalent thio­ether bond is established between reactive Cys57 and its flavin mononucleotide cofactor. This experiment validates the crystals to be suitable for in-depth follow-up studies of this still poorly understood signal-transduction mechanism. Importantly, the fixed-target delivery system also permitted a tenfold reduction in protein sample consumption compared with the more common high-viscosity extrusion-based delivery system. This development creates the prospect of an increase in XFEL project throughput for the field.




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The indexing ambiguity in serial femtosecond crystallography (SFX) resolved using an expectation maximization algorithm

An expectation maximization algorithm is implemented to resolve the indexing ambiguity which arises when merging data from many crystals in protein crystallography, especially in cases where partial reflections are recorded in serial femtosecond crystallography (SFX) at XFELs.




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Atmos & SFX Bundle by Resonance Sound on sale at 70% OFF

Plugin Boutique has launched an exclusive sale on the Atmos & SFX Bundle, featuring over 2,200 state-of-the-art SFX samples, loops and atmos by Sounds of Revolution and Audio Boutique. From robotic sound effects, complex impacts, mangled kick-free loops, mystic atmos and soundscapes, unique vocal fragments and much more, expand your sample collection with this unique […]

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SFXtools releases Sci-Fi Whooshes, Stutter & Noises and more

SFXtools has released its latest sound effects collection Sci-Fi Whooshes, a new “Film Audio” series pack featuring 64 high-quality unique sounding whoosh sound effects. Full of whoosh, swoosh, swish, passbys, flybys and movement sounds to spice up your sci-fi projects, trailers, cut scenes and movies. Every sound in the folder designed and delivered in industry-standard […]

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OC’s The Shakes Featured On StacksFX Blog

Sean Perry And Cameron Peinado Dish On Guitar Pedals In Recent Interview