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Related Experiment Video

Updated: Jun 21, 2026

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton
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Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton

Published on: February 25, 2021

Monolithic microfluidic mixing-spraying devices for time-resolved cryo-electron microscopy.

Zonghuan Lu1, Tanvir R Shaikh, David Barnard

  • 1Center for Integrated Electronics, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.

Journal of Structural Biology
|August 18, 2009
PubMed
Summary
This summary is machine-generated.

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A novel microfluidic device enables rapid mixing and spraying for time-resolved cryo-electron microscopy (cryo-EM). This method preserves the native structure of macromolecular complexes like ribosomes for structural analysis.

Area of Science:

  • Structural Biology
  • Biophysics
  • Microfluidics

Background:

  • Time-resolved cryo-electron microscopy (cryo-EM) aims to capture transient functional states of large biomolecules.
  • Rapid mixing, reaction, and vitrification are critical challenges in time-resolved cryo-EM.
  • Existing methods face limitations in achieving the necessary speed and sample quality.

Purpose of the Study:

  • To develop and validate a new methodology for time-resolved cryo-EM sample preparation.
  • To create a monolithic microfluidic device for integrated mixing, reaction, and sample deposition.
  • To assess the device's capability in preserving the structural integrity of macromolecular complexes.

Main Methods:

  • A single-chip microfabricated silicon device integrating a mixer, reaction channel, and pneumatic sprayer was designed.

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Last Updated: Jun 21, 2026

Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton
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  • Reactants were mixed using T-mixers and butterfly-shaped elements, reacted in a variable-length microfluidic channel.
  • Samples were sprayed as microdroplets onto an EM grid using humidified nitrogen gas, followed by plunge-freezing.

    • Main Results:

    • The device effectively mixed reactants and produced thin films suitable for cryo-EM within milliseconds.
    • Microdroplets spread optimally on hydrophilic carbon films.
    • Three-dimensional reconstructions of ribosomes showed preserved native structure, with subunits capable of reassociation.

    Conclusions:

    • The developed microfluidic device offers a robust solution for time-resolved cryo-EM sample preparation.
    • This methodology facilitates the study of dynamic structural changes in macromolecular complexes.
    • The device's performance indicates its potential for advancing structural biology research.