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

Abstract

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 inmore » 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.« less

Authors:
 [1];  [1]; ORCiD logo [1];  [1];  [1];  [2]; ORCiD logo [1];  [1];  [3]; ORCiD logo [3];  [3];  [3]; ORCiD logo [3]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [4];  [1];  [1]
  1. Arizona State Univ., Tempe, AZ (United States)
  2. Univ. of Gottingen (Germany)
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
  4. State Univ. of New York (SUNY), Buffalo, NY (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)
Sponsoring Org.:
National Science foundation (NSF); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES); Max-Planck Institute for Medical Research; CONACYT Fellowship
OSTI Identifier:
1605169
Grant/Contract Number:  
1231306; 1565180; R01GM095583; AC02-76SF00515
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Applied Crystallography (Online)
Additional Journal Information:
Journal Name: Journal of Applied Crystallography (Online); Journal Volume: 52; Journal Issue: 5; Journal ID: ISSN 1600-5767
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
47 OTHER INSTRUMENTATION; microfluidic devices; serial crystallography; injection; droplet generation

Citation Formats

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, and Ros, Alexandra. 3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating. United States: N. p., 2019. Web. https://doi.org/10.1107/s1600576719010343.
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. 3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating. United States. https://doi.org/10.1107/s1600576719010343
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, and Ros, Alexandra. Thu . "3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating". United States. https://doi.org/10.1107/s1600576719010343. https://www.osti.gov/servlets/purl/1605169.
@article{osti_1605169,
title = {3D printed droplet generation devices for serial femtosecond crystallography enabled by surface coating},
author = {Echelmeier, Austin and Kim, Daihyun and Cruz Villarreal, Jorvani and Coe, Jesse and Quintana, Sebastian and Brehm, Gerrit and Egatz-Gomez, Ana and Nazari, Reza and Sierra, Raymond G. and Koglin, Jason E. and Batyuk, Alexander and Hunter, Mark S. and Boutet, Sébastien and Zatsepin, Nadia and Kirian, Richard A. and Grant, Thomas D. and Fromme, Petra and Ros, Alexandra},
abstractNote = {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.},
doi = {10.1107/s1600576719010343},
journal = {Journal of Applied Crystallography (Online)},
number = 5,
volume = 52,
place = {United States},
year = {2019},
month = {8}
}

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    Works referencing / citing this record:

    Time-resolved serial femtosecond crystallography at the European XFEL
    journal, November 2019