Raster-scanning serial protein crystallography using micro- and nano-focused synchrotron beams

Abstract

A raster scanning serial protein crystallography approach is presented, that consumes as low ∼200–700 nl of sedimented crystals. New serial data pre-analysis software, NanoPeakCell, is introduced.

Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

“…The problem is further intensified when very small microcrystals are needed for efficient photoactivation in a pump-probe experiment. Often, radiation damage is avoided by collecting small portions of data from a larger number of crystals, for example, by using raster scanning in cryo-loops 32,34 or other fixed-target mounting devices 35 . Injectorbased serial crystallography pushes the principle to the extreme by exposing each crystal to X-rays only once, while providing a continuously refreshed supply of thousands of unexposed crystals.…”

Microfluidics: From crystallization to serial time-resolved crystallography

“…The resulting devices can then be sealed with $50 to 500 nm-thick silicon nitride windows. 34,46,[141][142][143][144] This approach allows for the creation of extremely small features, such as wells to capture individual microcrystals. However, the devices are relatively time and labor intensive and expensive to produce.…”

“…25,31,33,34,37,143,185,[191][192][193][194][195] The simplest mounting strategy involves the deposition of a crystal slurry between two thin silicon nitride wafers. 143 However, most approaches have utilized microfabricated mesh-type structures. One advantage of a meshtype structure is the potential to localize individual crystals within each small micro-well.…”

Protein Micro-Crystallography: Nanotechnology Challenges Ahead

“…The main-chain lysozyme structure was only slightly affected by radiation damage and there was practically no difference for the two beam sizes. The particularly high radiation dose for nanobeam SSX of 29 MGy/crystallite was tentatively attributed to a lag phase [20].The availability of up to KHz framing rate, zero readout noise pixel detectors -such as the EIGER detector from DECTRIS®-will allow further reducing protein consumption in SSX experiments.…”

“…This will favor in particular "brilliancehungry" experimental techniques, currently proposed only at few SR beamlines worldwide. Examples are sub-μm SR-MPX [19,20] (e.g. ID13 ESRF) ; ( Figure 2B) and time-resolved SR-MPX based on picoseconds X-ray flashes using an undulator harmonics ("pink" beam), a technique currently developed only for larger SR beam foci [21].…”