3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating

Echelmeier, Austin; Kim, Daihyun; Cruz Villarreal, Jorvani; Coe, Jesse; Quintana, Sebastian; Brehm, Gerrit; Egatz-Gomez, Ana; Nazari, Reza; Sierra, Raymond G.; Koglin, Jason E.; Batyuk, Alexander; Hunter, Mark S.; Boutet, Sébastien; Zatsepin, Nadia; Kirian, Richard A.; Grant, Thomas D.; Fromme, Petra; Ros, Alexandra

doi:10.1107/s1600576719010343

3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating

Journal Article · Thu Aug 29 00:00:00 EDT 2019 · Journal of Applied Crystallography (Online)

DOI:https://doi.org/10.1107/s1600576719010343· OSTI ID:1605169

Echelmeier, Austin ^[1]; Kim, Daihyun ^[1]; ^[1]; Coe, Jesse ^[1]; Quintana, Sebastian ^[1]; Brehm, Gerrit ^[2]; ^[1]; Nazari, Reza ^[1]; Sierra, Raymond G. ^[3]; ^[3]; Batyuk, Alexander ^[3]; Hunter, Mark S. ^[3]; ^[3]; ^[1]; ^[1]; ^[4]; Fromme, Petra ^[1]; Ros, Alexandra ^[1]

Arizona State Univ., Tempe, AZ (United States)
Univ. of Gottingen (Germany)
SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
State Univ. of New York (SUNY), Buffalo, NY (United States)

The role of surface wetting properties and their impact on the performance of 3D printed microfluidic droplet generation devices for serial femtosecond crystallography (SFX) are reported herein. SFX is a novel crystallography method enabling structure determination of proteins at room temperature with atomic resolution using X-ray free-electron lasers (XFELs). In SFX, protein crystals in their mother liquor are delivered and intersected with a pulsed X-ray beam using a liquid jet injector. Owing to the pulsed nature of the X-ray beam, liquid jets tend to waste the vast majority of injected crystals, which this work aims to overcome with the delivery of aqueous protein crystal suspension droplets segmented by an oil phase. For this purpose, 3D printed droplet generators that can be easily customized for a variety of XFEL measurements have been developed. The surface properties, in particular the wetting properties of the resist materials compatible with the employed two-photon printing technology, have so far not been characterized extensively, but are crucial for stable droplet generation. This work investigates experimentally the effectiveness and the long-term stability of three different surface treatments on photoresist films and glass as models for our 3D printed droplet generator and the fused silica capillaries employed in the other fluidic components of an SFX experiment. Finally, the droplet generation performance of an assembly consisting of the 3D printed device and fused silica capillaries is examined. Stable and reproducible droplet generation was achieved with a fluorinated surface coating which also allowed for robust downstream droplet delivery. Experimental XFEL diffraction data of crystals formed from the large membrane protein complex photosystem I demonstrate the full compatibility of the new injection method with very fragile membrane protein crystals and show that successful droplet generation of crystal-laden aqueous droplets intersected by an oil phase correlates with increased crystal hit rates.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory, Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)

Sponsoring Organization:: CONACYT Fellowship; Max-Planck Institute for Medical Research; National Institutes of Health (NIH); National Science foundation (NSF); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1605169

Journal Information:: Journal of Applied Crystallography (Online), Journal Name: Journal of Applied Crystallography (Online) Journal Issue: 5 Vol. 52; ISSN 1600-5767; ISSN JACGAR

Publisher:: International Union of CrystallographyCopyright Statement

Country of Publication:: United States

Language:: English

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