BEGIN:VCALENDAR VERSION:2.0 PRODID:-//CERN//INDICO//EN BEGIN:VEVENT SUMMARY:Current and future capabilities for serial and time resolved cryst allography at Diamond microfocus beamline VMXi DTSTART;VALUE=DATE-TIME:20250922T150000Z DTEND;VALUE=DATE-TIME:20250922T151500Z DTSTAMP;VALUE=DATE-TIME:20260526T060932Z UID:indico-contribution-1832@lindico453.srv.lu.se DESCRIPTION:Speakers: Juan Sanchez Weatherby (Diamond Light Source)\nThere is increasing interest in obtaining room temperature\, time resolved crys tal structures of\nproteins carrying out their biological functions. The t ransition between conventional cryogenic\nmacromolecular crystallography a nd serial crystallography involving microcrystals remains chal-\nlenging f or many projects. We have recently demonstrated the capability to measure good quality\,\nlow dose serial crystallography data from microcrystals wi thin crystallisation plates [1]. This capa-\nbility is available in the st andard operation mode of the beamline and does not require any specific\ns erial crystallography apparatus. In this approach\, microcrystals are tran sferred into a crystallisa-\ntion plate (typically 100 nL per drop) and ea ch droplet is subjected to raster scanning with a still\ndiffraction image measured every 10 µm. The resulting images are processed using standard serial\ndata processing software such as xia2.multiplex. This approach ena bles straightforward structure\ndetermination and analysis of crystal qual ity and unit cell parameters from non-optimised crys-\ntallisation conditi ons\, guiding users in their optimisation efforts. Very small quantities o f protein\nare required\, and the determination of a human peroxidase stru cture to 1.88 Angstrom resolution\nusing only 1.2 µL microcrystal suspens ion.\nSeveral approaches to sample delivery for time resolved crystallogra phy have been developed in-\ncluding droplet-on-demand tape drive-based sy stems developed for XFEL experiments that have\nbeen combined with X-ray e mission spectroscopy (XES) to monitor the redox and spin state of\nmetal - containing cofactors within the proteins [2]. However\, currently availabl e systems require\na large quantity of microcrystal sample as well as requ iring multiple skilled staff to operate. A\nnew system for serial crystall ography at VMXi is currently under development. This incorporates\na picol itre droplet-on-demand tape drive system capable of anaerobic operation to gether with an\nXES von Hamos spectrometer to enable spectroscopic validat ion in time resolved experiments of\nmetalloproteins. A compact design was required due to the tight spatial constraints of the VMXi\nend station th at was built for highly automated data collection from crystallisation pla tes\, and the\ndesign incorporates automation to reduce the number of pers onnel required to more closely ap-\nproach a typical synchrotron experimen t.\nProof of concept data obtained during the development process of the t ape drive and XES spec-\ntrometer will be presented\, including a high-res olution protein structure determined using the tape\ndrive system and XES data obtained from microcrystals of the copper enzyme nitrite reductase.\n [1] A.J. Thompson\, J. Sanchez-Weatherby\, L.J. Williams\, H. Mikolajek\, J. Sandy\, J.A.R. Worrall and\nM.A. Hough (2024) Efficient in situ screeni ng of and data collection from microcrystals in crystal-\nlization plates Acta Cryst.D80\, 279-288\n[2] Butyrin\, A. et al (2021) An on-demand\, dro p-on-drop method for studying enzyme catalysis by\nserial crystallography. Nature Methods 12\, 4461.\n\n\nAuthors:\nJuan Sanchez Weatherby\, Pierre Aller\, Amy Thompson\, Abby Telfer\, John Sutter\, James Sandy\, Halina Mi kolajek\, Matthew Rodrigues\, Mike Hough\, Allen Orville\n\nhttps://lindic o453.srv.lu.se/event/583/contributions/1832/ LOCATION:LINXS at The Loop URL:https://lindico453.srv.lu.se/event/583/contributions/1832/ END:VEVENT END:VCALENDAR