Plug-and-play polymer microfluidic chips for hydrated, room temperature, fixed-target serial crystallography

Deepshika Gilbile, Megan L. Shelby, Artem Y. Lyubimov, Jennifer L. Wierman, Diana C.F. Monteiro, Aina E. Cohen, Silvia Russi, Matthew A. Coleman, Matthias Frank, Tonya L. Kuhl

Research output: Contribution to journalArticlepeer-review

Abstract

The practice of serial X-ray crystallography (SX) depends on efficient, continuous delivery of hydrated protein crystals while minimizing background scattering. Of the two major types of sample delivery devices, fixed-target devices offer several advantages over widely adopted jet injectors, including: lower sample consumption, clog-free delivery, and the ability to control on-chip crystal density to improve hit rates. Here we present our development of versatile, inexpensive, and robust polymer microfluidic chips for routine and reliable room temperature serial measurements at both synchrotrons and X-ray free electron lasers (XFELs). Our design includes highly X-ray-transparent enclosing thin film layers tuned to minimize scatter background, adaptable sample flow layers tuned to match crystal size, and a large sample area compatible with both raster scanning and rotation based serial data collection. The optically transparent chips can be used both for in situ protein crystallization (to eliminate crystal handling) or crystal slurry loading, with prepared samples stable for weeks in a humidified environment and for several hours in ambient conditions. Serial oscillation crystallography, using a multi-crystal rotational data collection approach, at a microfocus synchrotron beamline (SSRL, beamline 12-1) was used to benchmark the performance of the chips. High-resolution structures (1.3-2.7 Å) were collected from five different proteins-hen egg white lysozyme, thaumatin, bovine liver catalase, concanavalin-A (type VI), and SARS-CoV-2 nonstructural protein NSP5. Overall, our modular fabrication approach enables precise control over the cross-section of materials in the X-ray beam path and facilitates chip adaption to different sample and beamline requirements for user-friendly, straightforward diffraction measurements at room temperature.

Original languageEnglish (US)
Pages (from-to)4831-4845
Number of pages15
JournalLab on a Chip
Volume21
Issue number24
DOIs
StatePublished - Dec 21 2021
Externally publishedYes

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Chemistry(all)
  • Biomedical Engineering

Fingerprint

Dive into the research topics of 'Plug-and-play polymer microfluidic chips for hydrated, room temperature, fixed-target serial crystallography'. Together they form a unique fingerprint.

Cite this