Plug-and-play polymer microfluidic chips for hydrated, room-temperature fixed-target serial crystallography
| Autor: | Deepshika Gilbile, Matthew A. Coleman, Artem Y. Lyubimov, Megan L. Shelby, Aina E. Cohen, Tonya L. Kuhl, Silvia Russi, Diana C. F. Monteiro, Jennifer L. Wierman, Matthias Frank |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Materials science
Fabrication Serial communication Polymers Microfluidics Biomedical Engineering Bioengineering 02 engineering and technology Crystallography X-Ray Biochemistry Article law.invention Crystal 03 medical and health sciences law Animals Humans 030304 developmental biology 0303 health sciences SARS-CoV-2 Temperature COVID-19 General Chemistry Equipment Design 021001 nanoscience & nanotechnology Synchrotron Crystallography Beamline Cattle 0210 nano-technology Raster scan Protein crystallization |
| Zdroj: | Lab Chip |
| Popis: | 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 A) 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. |
| Databáze: | OpenAIRE |
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