Enabling Structural Biology at Physiological Temperatures - New Tools for Sample Preparation, Transport and Data Collection at Controlled Humidity

We will develop robust tools that enable fully automated crystallography experiments at synchrotron beamlines using samples at ambient temperatures and under controlled humidity conditions These tools include new equipment for sample preparation, for sample transport and for robotic sample mounting onto a beamline goniometer The proposed phase 1 project will provide robust commercializable tools and prove the technical feasibility of our approach for sample preparation and shipping In addition, we will develop a design for a humidity-controlled sample transport chamber that is compatible with the majority of robotic sample exchange systems used at crystallography beamlines worldwide, including at the ALS, NSLS and APS As Light Source facilities worldwide are initiating serial crystallography programs and acquiring fast frame-rate area detectors, more experiments will be done using samples at ambient temperatures enabling new approaches to study protein dynamics There however is not yet a reliable way to store and ship crystalline samples to the synchrotron on holders compatible with these experiments Our developments will expand what is now only possible using samples stored at cryogenic temperatures, to enable fully automated and remote-access crystallography under controlled humidity conditions This proposal takes advantage of tools developed by the SSRL-SMB group at Stanford that include a prototype plate useful for sample crystallization, sample transport and robotic sample mounting using a sample exchange robot all under humidity-controlled conditions A first aim of this phase-1 project is to develop a commercializable process for the production of the SMB-developed plates and associated tools To enhance the usability of these plates, an important part of this aim is to develop a reliable temperature controlled (+/- 25ᵒC for 7 days) shipping container and spill-proof inserts for the plates A second aim will expand the utility of this plate to all sample mounting robots used for crystallography experiments at synchrotrons in the US, by developing a design for an automated humidity-controlled transport chamber that can remove sample pins from the plate and present them in the orientation compatible with a variety of robotic grippers