Mixing injector enables time-resolved crystallography with high hit rate at X-ray free electron lasers.
Knowledge of protein structure provides essential insight into function, enhancing our understanding of diseases and enabling new treatment development. X-ray crystallography has been used to solve the structures of more than 100 000 proteins; however, the vast majority represent long-lived states that do not capture the functional motions of these molecular machines. Reactions triggered by the addition of a ligand can be the most challenging to detect with crystallography because of the difficulty of synchronizing reactions to create detectable quantities of transient states. The development of X-ray free electron lasers (XFELs) and serial femtosecond crystallography (SFX) enables new approaches for solving protein structures following the rapid diffusion of ligands into micron sized protein crystals. Conformational changes occurring on millisecond timescales can be detected and time-resolved. Here, we describe a new XFEL injector which incorporates a microfluidic mixer to rapidly combine reactant and sample milliseconds before the sample reaches the X-ray beam. The mixing injector consists of bonded, concentric glass capillaries. The fabrication process, employing custom laser cut centering spacers and UV curable epoxy, ensures precise alignment of capillaries for repeatable, centered sample flow and dependable mixing. Crystal delivery capillaries are 50 or 75 μm in diameter and can contain an integrated filter depending on the demands of the experiment. Reaction times can be varied from submillisecond to several hundred milliseconds. The injector features rapid and uniform mixing, low sample dilution, and high hit rates. It is fully compatible with existing SFX beamlines.
School of Applied and Engineering Physics, Cornell University , Ithaca, New York 14853, USA.
George D
GD
Calvey
School of Applied and Engineering Physics, Cornell University , Ithaca, New York 14853, USA.
Andrea M
AM
Katz
Meinig School of Biomedical Engineering, Cornell University , Ithaca, New York 14853, USA.
Chris B
CB
Schaffer
School of Applied and Engineering Physics, Cornell University , Ithaca, New York 14853, USA.
Lois
L
Pollack
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