Abstract
Serial femtosecond crystallography (SFX) with femtosecond X-ray pulses from X-ray freeelectron lasers (XFELs) offers a route to overcome radiation damage to small protein crystals
via the 'diffraction-before-destruction' approach. A single-pulse X-ray exposure completely
destroys individual crystals; therefore, fresh specimens must be supplied for subsequent X-ray
pulses to continue data acquisition. A matrix method using protein crystal carriers, such as
grease and oil-free hydrogel media, has been developed for serial sample loading. The microextrusion technique employing matrices allows the structure determination of a wide variety of
proteins with low sample consumption, typically less than 1 mg. This offers new opportunities
for time-resolved studies of light-driven structural changes and chemical dynamics using pumpprobe techniques at XFEL and synchrotron facilities.