Title: A user-friendly plug-and-play cyclic olefin copolymer-based microfluidic chip for room-temperature, fixed-target serial crystallography

Abstract

Over the past two decades, serial X-ray crystallography has enabled the structure determination of a wide range of proteins. With the advent of X-ray free-electron lasers (XFELs), ever-smaller crystals have yielded high-resolution diffraction and structure determination. A crucial need to continue advancement is the efficient delivery of fragile and micrometre-sized crystals to the X-ray beam intersection. This paper presents an improved design of an all-polymer microfluidic `chip' for room-temperature fixed-target serial crystallography that can be tailored to broadly meet the needs of users at either synchrotron or XFEL light sources. The chips are designed to be customized around different types of crystals and offer users a friendly, quick, convenient, ultra-low-cost and robust sample-delivery platform. Compared with the previous iteration of the chip [Gilbile et al. (2021), Lab Chip , 21 , 4831–4845], the new design eliminates cleanroom fabrication. It has a larger imaging area to volume, while maintaining crystal hydration stability for both in situ crystallization or direct crystal slurry loading. Crystals of two model proteins, lysozyme and thaumatin, were used to validate the effectiveness of the design at both synchrotron (lysozyme and thaumatin) and XFEL (lysozyme only) facilities, yielding complete data sets with resolutions of 1.42, 1.48 and 1.70 Å,more » respectively. Overall, the improved chip design, ease of fabrication and high modifiability create a powerful, all-around sample-delivery tool that structural biologists can quickly adopt, especially in cases of limited sample volume and small, fragile crystals.« less

Authors:

Liu, Zhongrui; Gu, Kevin K.; Shelby, Megan L. ;

• Search DOE PAGES for author "Shelby, Megan L."
• Search DOE PAGES for ORCID "0000-0001-6984-957X"

Gilbile, Deepshika; Lyubimov, Artem Y.; Russi, Silvia ;

• Search DOE PAGES for author "Russi, Silvia"
• Search DOE PAGES for ORCID "0000-0002-9666-1465"

Cohen, Aina E. ;

• Search DOE PAGES for author "Cohen, Aina E."
• Search DOE PAGES for ORCID "0000-0003-2414-9427"

Narayanasamy, Sankar Raju; Botha, Sabine; Kupitz, Christopher; Sierra, Raymond G.; Poitevin, Fredric; Gilardi, Antonio; Lisova, Stella; Coleman, Matthew A.; Frank, Matthias; Kuhl, Tonya L.

• Search DOE PAGES for author "Kuhl, Tonya L."
• Search DOE PAGES for ORCID "0000-0002-6069-0205"

Publication Date:

Mon Sep 25 00:00:00 EDT 2023

Research Org.:

Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Org.:

USDOE National Nuclear Security Administration (NNSA); National Science Foundation (NSF); National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:

2008115

Alternate Identifier(s):

OSTI ID: 2229586

Report Number(s):

LLNL-JRNL-844105
Journal ID: ISSN 2059-7983; ACSDAD; PII: S2059798323007027

Grant/Contract Number:  

AC52-07NA27344; 1231306; R01GM117342; U19 AI144184; AC02-76SF00515; P30GM133894

Resource Type:

Published Article

Journal Name:

Acta Crystallographica. Section D. Structural Biology

Additional Journal Information:

Journal Name: Acta Crystallographica. Section D. Structural Biology Journal Volume: 79 Journal Issue: 10; Journal ID: ISSN 2059-7983

Publisher:

International Union of Crystallography (IUCr)

Country of Publication:

United Kingdom

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; x-ray crystallography; synchrotrons; XFELS; sample delivery; fixed targets; cyclic olefin copolymer; microfluidics; serial crystallography; room-temperature SFX