An integrated high-throughput data acquisition system for biological solution X-ray scattering studies - PubMed (original) (raw)
An integrated high-throughput data acquisition system for biological solution X-ray scattering studies
Anne Martel et al. J Synchrotron Radiat. 2012 May.
Abstract
A fully automated high-throughput solution X-ray scattering data collection system has been developed for protein structure studies at beamline 4-2 of the Stanford Synchrotron Radiation Lightsource. It is composed of a thin-wall quartz capillary cell, a syringe needle assembly on an XYZ positioning arm for sample delivery, a water-cooled sample rack and a computer-controlled fluid dispenser. It is controlled by a specifically developed software component built into the standard beamline control program Blu-Ice/DCS. The integrated system is intuitive and very simple to use, and enables experimenters to customize data collection strategy in a timely fashion in concert with an automated data processing program. The system also allows spectrophotometric determination of protein concentration for each sample aliquot in the beam via an in situ UV absorption spectrometer. A single set of solution scattering measurements requires a 20-30 µl sample aliquot and takes typically 3.5 min, including an extensive capillary cleaning cycle. Over 98.5% of measurements are valid and free from artefacts commonly caused by air-bubble contamination. The sample changer, which is compact and light, facilitates effortless switching with other sample-handling devices required for other types of non-crystalline X-ray scattering experiments.
Figures
Figure 1
Schematic view of the key hardware components of the sample changer (a). It consists of a motorized XYZ translation arm (A), a syringe needle (B), a temperature-controlled sample rack in the 96-well microplate format (C), a temperature-controlled capillary holder assembly (D) and a quartz capillary. The capillary holder assembly (D), made of anodized aluminium, interfaces the capillary to compressed air via a valve for drying, as well as two syringes, for capillary cleaning and sample aliquot handling. The gentle compression of the syringe needle holder onto the capillary holder assembly makes an air-tight O-ring seal, enabling precise positional control of the sample aliquot within the quartz capillary. The inset (b) depicts the capillary holder assembly in detail. It consists of three parts: an air valve (D1) that switches the compressed air flow for drying the capillary, an interface part (D2) with internal channels (as outlined in the drawing) connecting the capillary to the cleaning syringe, the air valve and the syringe needle, and a capillary holder (D3) into which the quartz capillary is glued. The O-rings sealing the connections between (D2) and (D3) as well as between (D2) and the syringe needle holder are also depicted. Samples are placed on the sample rack in two different patterns (c) for dilution series (columns 1 and 2) or for titration series (columns 4 and 5): the white wells represent buffer solutions, the coloured wells protein samples and the blue wells empty.
Figure 2
Photograph of the sample changer hardware. Depicted are the motorized XYZ translation arm (A), the syringe needle (B), the temperature-controlled sample rack (C), and the temperature-controlled capillary holder assembly (D). The X-ray capillary is housed within a protective aluminium sheath (E) with two pairs of X-ray and UV/vis ports, perpendicular to the capillary long axis. A piece of plastic tubing is connected to the bottom opening of the capillary for drain collection (not depicted).
Figure 3
The SolSAXS Blu-Ice tab showing a set of sample handling and exposure command sequences (A) and global data acquisition parameters (B). A separate set of data acquisition parameters, for example frequency of blank (buffer) measurement and number/length of X-ray exposures, can be defined for each column of the sample rack (C).
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