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On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature
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Room-temperature macromolecular crystallography using a micro-patterned silicon chip with minimal background

Philip Roedig1, Ramona Duman2, Juan Sanchez-Weatherby2

  • 1Deutsches Elektronen-Synchrotron DESY, Photon Science, Notkestrasse 85, Hamburg 22607, Germany.

Journal of Applied Crystallography
|June 9, 2016
PubMed
Summary

A novel silicon chip enables faster serial crystallography by holding thousands of microcrystals. This method allows room-temperature data collection, significantly reducing radiation damage effects compared to cryogenic methods.

Keywords:
X-ray radiation damagecrystallography on a chiproom-temperature crystallographysynchrotron serial crystallography

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Area of Science:

  • Structural Biology
  • Crystallography
  • Materials Science

Background:

  • Serial crystallography experiments are gaining traction at synchrotron sources, driven by advances in X-ray free-electron lasers.
  • High-speed detectors (up to 1 kHz) necessitate rapid sample exchange, which has become a bottleneck in these experiments.
  • Existing methods face challenges with crystal lifetime (milliseconds) and sample handling.

Purpose of the Study:

  • To develop an efficient sample holder for serial crystallography experiments.
  • To overcome the sample exchange bottleneck in high-repetition-rate X-ray diffraction.
  • To enable room-temperature data collection with reduced radiation damage.

Main Methods:

  • Development of a micro-patterned chip from single-crystalline silicon for holding microcrystals.
  • Loading of microcrystals onto the chip with efficient removal of excess mother liquor.
  • Room-temperature data collection of insulin crystals using humidified air and subsequent structure refinement.

Main Results:

  • The silicon chip successfully holds thousands of microcrystals with minimal background.
  • Room-temperature data collection from insulin crystals demonstrated the chip's utility in macromolecular crystallography.
  • Insulin crystals showed no radiation-damage-induced structural changes up to 565.6 kGy, with half-decay dose (D1/2) of 147.5 ± 19.1 kGy at room temperature.

Conclusions:

  • The micro-patterned silicon chip effectively addresses the sample exchange bottleneck in serial crystallography.
  • Room-temperature data collection using this chip significantly enhances crystal radiation resistance compared to cryogenic conditions.
  • This approach facilitates high-throughput structural biology studies at synchrotron sources.