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Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton
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Spitrobot-2 advances time-resolved cryo-trapping crystallography to under 25 ms.

Maria Spiliopoulou1, Caitlin E Hatton2, Martin Kollewe3

  • 1University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.

Communications Chemistry
|November 20, 2025
PubMed
Summary

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The new spitrobot-2 system enhances cryo-trapping time-resolved crystallography with millisecond resolution. This faster, user-friendly device enables new insights into protein dynamics and ligand binding for structural biology research.

Area of Science:

  • Structural Biology
  • Biophysics
  • Biochemistry

Background:

  • The spitrobot system previously enabled millisecond time-resolved crystallography using cryo-trapping.
  • Advancements were needed to improve user-friendliness and reduce cryo-trapping delay times.

Purpose of the Study:

  • To introduce spitrobot-2, an improved integrated benchtop device for enhanced cryo-trapping.
  • To demonstrate the capabilities of spitrobot-2 in capturing fast biological processes.

Main Methods:

  • Development of an integrated, user-friendly benchtop device (spitrobot-2).
  • Implementation of semi-automatic sample exchange and an automated shutter.
  • Reduction of cryo-trapping delay time to 23 milliseconds.

Main Results:

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  • Spitrobot-2 achieves a cryo-trapping delay time of 23 ms, twice as fast as the previous generation.
  • Successful cryo-trapping was demonstrated within 25 ms, capturing conformational changes and ligand binding.
  • 12 crystal structures from three model systems confirmed the system's efficacy.

Conclusions:

  • Spitrobot-2 significantly enhances access to time-resolved X-ray crystallography.
  • The improved system empowers the structural biology community with advanced tools for research.
  • Faster cryo-trapping expands the scope of target systems addressable by this technique.