Protein Micro-Crystallography: Nanotechnology Challenges Ahead

Abstract

The status and prospects of protein microcrystallography (MPX) at high brilliance synchrotron radiation sources are reviewed. We discuss emerging trends in miniaturizing sample environments for serial crystallography (SX) experiments allowing manipulation and positioning of biological objects down to nanoscale dimensions with low contact forces.

Perspectives: DOD Inkjets at High and Ultra-Brilliant Light Sources

“…Inter-pulse distances and pulsetrains can be changed according to specific filling patterns. The strongest undulator harmonics will provide a flux of ~10 10 photons/pulse for a Δλ/λ~2.8*10 -2 band-width (pink beam) [25,26], enabling single pulse scattering with μm to sub-μm spots for the size of droplets shown in figure 2A-2C. Scaling with the volume suggests that droplets of d = 8 μm (268 aL) for EBS/ESRF and d = 1.7 μm (2.5 aL) for EuXFEL should show equivalent scattering with a single pulse as d = 80 μm droplets (268 pL) at the ESRF-ID13 beamline ( Figure 2B and 2C).…”

Microfluidics: From crystallization to serial time-resolved crystallography

“…Finally, we discuss the potential for these various microfluidic strategies to address future challenges related to the study of protein structures and dynamics. As this review is focused on the technologies surrounding crystal growth and delivery, and not on the larger aspects of serial [43][44][45][46] or time-resolved X-ray methods, [46][47][48] we refer the reader to other excellent reviews on these topics.…”

Graphene-based microfluidics for serial crystallography

“…1–4 One goal of this micro-focused technology is to enhance the diffraction signal from micro-crystals by matching the size of the beam to the size of the crystal. Here, the signal enhancement comes from a reduction in the level of background scatter associated with the solvent or mounting support surrounding the crystal.…”

“…10,11,15,18–45 However, this data collection strategy suffers from the need to grow and efficiently manipulate a large number of fragile micro-crystals. Reported methods include the mounting of many crystals using traditional, loop-style holders, 38,46 a variety of fixed-target architectures, 4,6,11,47–55 and different micro-crystal injection strategies 10,52,56–63 However, nearly all of these strategies have focused on transferring a pre-prepared slurry of micro-crystals onto a mount or into an injector, rather than coupling the processes of crystallization and X-ray analysis. 6,10,47,48,62 This requirement for physical handling introduces the need for additional experimental procedures, can potentially damage fragile or sensitive crystals, and particularly in the case of micro-crystal injection jets, can lead to inefficient sample utilization 6,10,47,48,62 …”

“…The application of micro-crystallography in structural biology has accelerated in recent years due to technological advances that enable the use of smaller and ever more brilliant X-ray beams. [1][2][3][4] One goal of this micro-focused technology is to enhance the diffraction signal from micro-crystals by matching the size of the beam to the size of the crystal. Here, the signal enhancement comes from a reduction in the level of background scatter associated with the solvent or mounting support surrounding the crystal.…”