Abstract
ID23-2 is a fixed-energy (14.2 keV) microfocus beamline at the European Synchrotron Radiation Facility (ESRF) dedicated to macromolecular crystallography. The optics and sample environment have recently been redesigned and rebuilt to take full advantage of the upgrade of the ESRF to the fourth generation Extremely Brilliant Source (ESRF-EBS). The upgraded beamline now makes use of two sets of compound refractive lenses and multilayer mirrors to obtain a highly intense (>1013 photons s-1) focused microbeam (minimum size 1.5 µm × 3 µm full width at half-maximum). The sample environment now includes a FLEX-HCD sample changer/storage system, as well as a state-of-the-art MD3Up high-precision multi-axis diffractometer. Automatic data reduction and analysis are also provided for more advanced protocols such as synchrotron serial crystallographic experiments.
Highlights
Macromolecular microcrystallography is an indispensable tool in modern structural biology
Pioneering studies in macromolecular crystallography (MX) using microbeams were performed on microfocus beamlines such as European Synchrotron Radiation Facility (ESRF) ID13 (Cusack et al, 1998) which laid the groundwork for many of the techniques and instrumentation currently employed today (Perrakis et al, 1999)
There were no microfocus beamlines dedicated to MX, which was the motivation for the design and construction of the first ‘dedicated’ macromolecular microcrystallography beamline: ID23-2 (Flot et al, 2010)
Summary
Macromolecular microcrystallography is an indispensable tool in modern structural biology. Small beams facilitate more advanced data collection strategies in which multiple, usually low-completeness, subdatasets are collected from multiple microcrystals and/or positions on non-microcrystals by rapidly scanning across sample holders and simultaneously collecting data at high frame rates (Liu et al, 2013; Gati et al, 2014; Stellato et al, 2014; Nogly et al, 2015) These so-called ‘serial crystallography’ experiments can be performed on a wide variety of samples in diverse sample supports and across a wide range of crystal sizes at both cryo and ambient temperatures. All of these aspects of MX microcrystallography, concomitantly with the design of the ESRF-EBS [Extremely Brilliant Source (Raimondi, 2016)] with its dramatic improvement in brilliance, were the motivations for upgrading the ID23-2 beamline
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