Speaker
Description
Serial macromolecular crystallography (SMX) has become a powerful method for resolving
the structures of biological macromolecules at room temperature (RT). Although microfocus
beamlines at third-generation synchrotrons are instrumental, their data acquisition is typically
limited to the millisecond scale due to constraints in photon flux and detector speed. The
newly developed ID29 beamline at the European Synchrotron Radiation Facility (ESRF)—a
flagship of the Extremely Brilliant Source (EBS) upgrade—was purpose-built to leverage the
capabilities of this fourth-generation source. As the first beamline dedicated to
room-temperature serial microsecond crystallography (RT-SµX) with true microsecond X-ray
pulses, ID29 features a compact, flexible diffractometer that supports rapid sample exchange
and accommodates multiple solid supports and three types of high-viscosity extruders
(HVEs). Our study highlights the critical integration of pulsed beams, the fast JungFrau4M
detector, and synchronized data acquisition systems for effective RT-SµX experiments. The
unique beam properties of the new ESRF source enable microsecond time-resolved
crystallography, yielding high-quality electron density maps from relatively few merged
frames. These advances position RT-SµX at ID29 as a model for future applications at
upcoming fourth-generation synchrotron facilities worldwide.
