FMX – the Frontier Microfocusing Macromolecular Crystallography Beamline at the National Synchrotron Light Source II

Abstract

Two new macromolecular crystallography (MX) beamlines at the National Synchrotron Light Source II, FMX and AMX, opened for general user operation in February 2017 [Schneider et al. (2013). J. Phys. Conf. Ser. 425, 012003; Fuchs et al. (2014). J. Phys. Conf. Ser. 493, 012021; Fuchs et al. (2016). AIP Conf. Proc. SRI2015, 1741, 030006]. FMX, the micro-focusing Frontier MX beamline in sector 17-ID-2 at NSLS-II, covers a 5–30 keV photon energy range and delivers a flux of 4.0 × 1012 photons s−1 at 1 Å into a 1 µm × 1.5 µm to 10 µm × 10 µm (V × H) variable focus, expected to reach 5 × 1012 photons s−1 at final storage-ring current. This flux density surpasses most MX beamlines by nearly two orders of magnitude. The high brightness and microbeam capability of FMX are focused on solving difficult crystallographic challenges. The beamline's flexible design supports a wide range of structure determination methods – serial crystallography on micrometre-sized crystals, raster optimization of diffraction from inhomogeneous crystals, high-resolution data collection from large-unit-cell crystals, room-temperature data collection for crystals that are difficult to freeze and for studying conformational dynamics, and fully automated data collection for sample-screening and ligand-binding studies. FMX's high dose rate reduces data collection times for applications like serial crystallography to minutes rather than hours. With associated sample lifetimes as short as a few milliseconds, new rapid sample-delivery methods have been implemented, such as an ultra-high-speed high-precision piezo scanner goniometer [Gao et al. (2018). J. Synchrotron Rad. 25, 1362–1370], new microcrystal-optimized micromesh well sample holders [Guo et al. (2018). IUCrJ, 5, 238–246] and highly viscous media injectors [Weierstall et al. (2014). Nat. Commun. 5, 3309]. The new beamline pushes the frontier of synchrotron crystallography and enables users to determine structures from difficult-to-crystallize targets like membrane proteins, using previously intractable crystals of a few micrometres in size, and to obtain quality structures from irregular larger crystals.

The temperature-dependent conformational ensemble of SARS-CoV-2 main protease (Mpro)

“…X-ray data were collected at the National Synchrotron Light Source II (NSLS-II) beamline 17-ID-2 (FMX) (Schneider et al, 2021) using an X-ray beam of energy 12.66 keV, corresponding to a wavelength of 0.9793 Å; a horizontal-bounce Si111 double crystal monochromator; and an Eiger X 16M pixel array detector (Dectris). Temperature at the sample goniometer was controlled using a Cryostream 800 (OxfordCryosystems).…”

Robotic sample changers for macromolecular X-ray crystallography and biological small-angle X-ray scattering at the National Synchrotron Light Source II

“…The implementation of the Jlib system, from the robot task layer written in the VAL3 language (Stä ubli, Switzerland), up to the included 'EMBL' EPICS-based input-output controller (IOC) have been implemented as described by Papp, Felisaz et al (2017). Both the LiX Bluesky server (https://nsls-ii.github.io/bluesky/) and AMX/FMX Life Science Data Collection (LSDC) server (Schneider et al, 2021) interface with the robot control system via a Python (Van Rossum & Drake, 2009) based robot control library customized for each beamline, which provides both control of and feedback from the robot system.…”

“…The Advanced Beamlines for Biological Investigations using X-rays project, which was funded by the National Institutes of Health, constructed two MX beamlines and one bioSAXS beamline: the highly Automated Macromolecular Crystallography beamline (AMX; https://www.bnl. gov/nsls2/beamlines/beamline.php?r=17-ID-1), the Frontier Macromolecular Crystallography beamline (FMX; Schneider et al, 2021) and the Life Science X-ray scattering beamline (LiX; Yang et al, 2020). Due to the brightness of the National Synchrotron Light Source II (NSLS-II), these beamlines can achieve fluxes greater than 10 12 photons s À1 at the sample.…”

AlphaFold Protein Structure Database for Sequence-Independent Molecular Replacement

“…Diffraction data were collected at the NSLS-II beamline FMX (17ID-2) at 100 K (Fuchs et al ., 2016, Schneider et al ., 2021). The beamline is equipped with an Eiger 16M detector.…”