Megahertz pulse trains enable multi-hit serial femtosecond crystallography experiments at X-ray free electron lasers

The European X-ray Free Electron Laser (XFEL) and Linac Coherent Light Source (LCLS) II are extremely intense sources of X-rays capable of generating Serial Femtosecond Crystallography (SFX) data at megahertz (MHz) repetition rates. Previous work has shown that it is possible to use consecutive X-ray pulses to collect diffraction patterns from individual crystals. Here, we exploit the MHz pulse structure of the European XFEL to obtain two complete datasets from the same lysozyme crystal, first hit and the second hit, before it exits the beam. The two datasets, separated by <1 µs, yield up to 2.1 Å resolution structures. Comparisons between the two structures reveal no indications of radiation damage or significant changes within the active site, consistent with the calculated dose estimates. This demonstrates MHz SFX can be used as a tool for tracking sub-microsecond structural changes in individual single crystals, a technique we refer to as multi-hit SFX

Time-Resolved Analysis of the Structural Dynamics of Assembling Gold Nanoparticles

The hydrophobic collapse is a structural transition of grafted polymer chains in a poor solvent. Although such a transition seems an intrinsic event during clustering of polymer-stabilized nanoparticles in the liquid phase, it has not been resolved in real time. In this work, we implemented a microfluidic 3D-flow-focusing mixing reactor equipped with real-time analytics, small-angle X-ray scattering (SAXS), and UV-vis-NIR spectroscopy to study the early stage of cluster formation for polystyrene-stabilized gold nanoparticles. The polymer shell dynamics obtained by in situ SAXS analysis and numerical simulation of the solvent composition allowed us to map the interaction energy between the particles at early state of solvent mixing, 30 ms behind the crossing point. We found that the rate of hydrophobic collapse depends on water concentration, ranging between 100 and 500 nm/s. Importantly, we confirmed that the polymer shell collapses prior to the commencement of clustering