Automated Sample Mounting and Alignment System for Biological Crystallography at a Synchrotron Source

Abstract

High-throughput data collection for macromolecular crystallography requires an automated sample mounting and alignment system for cryo-protected crystals that functions reliably when integrated into protein-crystallography beamlines at synchrotrons. Rapid mounting and dismounting of the samples increases the efficiency of the crystal screening and data collection processes, where many crystals can be tested for the quality of diffraction. The sample-mounting subsystem has random access to 112 samples, stored under liquid nitrogen. Results of extensive tests regarding the performance and reliability of the system are presented. To further increase throughput, we have also developed a sample transport/storage system based on "puck-shaped" cassettes, which can hold sixteen samples each. Seven cassettes fit into a standard dry shipping Dewar. The capabilities of a robotic crystal mounting and alignment system with instrumentation control software and a relational database allows for automated screening and data collection to be developed.

Operator-assisted harvesting of protein crystals using a universal micromanipulation robot

“…Computer-assisted robotic crystallization pipelines are capable of cocktail preparation, crystallization plate setup, and inspection and interpretation of results. The mounting of crystal pins, data collection and structure solution are also highly automated (Karain et al, 2002;Pflugrath, 2004;Snell et al, 2004;Beteva et al, 2006), and more and more beamlines are being equipped with pin mounting robotics (http://smb.slac.stanford.edu/robosync/). Numerous pharmaceutical companies have thus implemented powerful crystallography pipelines, capable of screening numerous drug targets and receptor-ligand complexes per day (Blundell et al, 2002;Hosfield et al, 2003;Congreve et al, 2006).…”

Automation, parallelism, and robotics for proteomics

“…Automated robot used to mount and align protein crystals at Berkeley Lab Advanced Light Source. Reproduced, with permission, from Thomas Earnest (LBL) [40].…”

Recent improvements to the automatic characterization and data collection algorithms on MASSIF-1

“…Advances in robotics and software have been key in these developments and have had a particular impact on structural biology, allowing multiple constructs to be screened and purified (Camper & Viola, 2009;Hart & Waldo, 2013;Vijayachandran et al, 2011); huge numbers of crystallisation experiments to be performed (Elsliger et al, 2010;Ferrer et al, 2013;Heinemann et al, 2003;Joachimiak, 2009;Calero et al, 2014), samples to be mounted at synchrotrons (Cipriani et al, 2006;Cohen et al, 2002;Jacquamet et al, 2009;Papp et al, 2017;Snell et al, 2004), data to be analysed and processed (Bourenkov & Popov, 2010;Holton & Alber, 2004;Incardona et al, 2009;Leslie et al, 2002;Monaco et al, 2013;Winter, 2010) and the entire PDB to be validated (Joosten et al, 2012). The combination of robotic sample mounting and on-line data analysis has been particularly important in macromolecular crystallography (MX) as it allowed time to be saved, large numbers of samples to be screened and enabled the remote operation of beamlines.…”