Berlin 2018 – scientific programme

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KFM: Fachverband Kristalline Festkörper und deren Mikrostruktur

KFM 23: Lithography III: Lithography and Structuring (joint session KFM/DS)

KFM 23.2: Talk

Thursday, March 15, 2018, 10:00–10:20, EMH 025

3D printing at the diffraction limit: sample injection for time-resolved serial crystallography — •Michael Heymann — MPI of Biochemistry, Am Klopferspitz 18, 82152 Martinsried

Continuous injection using the Gas Dynamic Virtual Nozzle (GDVN) is a proven sample delivery method for biological imaging using X-ray free-electron lasers. However, many important aspects of GDVN functionality have yet to be thoroughly understood and/or refined due to fabrication limitations. We report the application of 2-photon polymerization as a form of high-resolution 3D printing to fabricate GDVNs with submicron resolution. This allows rapid prototyping of a wide range of nozzle designs from standard CAD drawings to iteratively optimize crucial dimensions for optimal performance. To understand enzyme catalysis and protein conformational changes at the atomic scale, we pioneered mixing-injectors for time-resolved structural biology to record molecular movies of substrate turn-over. We experimentally validate 3D print accuracy, as well as fluid mixing dynamics using X-ray tomographic imaging. We developed mixing-injectors to mix nanocrystals with substrate and to subsequently deliver them into the X-ray interaction region just milliseconds after mixing. This method can determine the structures of transient states and thereby kinetic mechanisms. In a proof of principle experiment, we could follow the catalytic reaction of the M. tuberculosis β-lactamase with the 3rd generation antibiotic ceftriaxone by time-resolved serial crystallography with millisecond to second time resolution at 2Å spatial resolution.