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Related Concept Videos

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

3.7K
Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
3.7K

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Related Experiment Video

Updated: Oct 12, 2025

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy
10:05

Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy

Published on: November 6, 2021

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Fast Grid Preparation for Time-Resolved Cryo-Electron Microscopy.

David P Klebl1, Frank Sobott2, Howard D White3

  • 1School of Biomedical Sciences, Faculty of Biological Sciences & Astbury Centre for Structural and Molecular Biology, University of Leeds.

Journal of Visualized Experiments : Jove
|November 22, 2021
PubMed
Summary
This summary is machine-generated.

Rapid advancements in cryo-electron microscopy (cryo-EM) enable faster grid preparation using new methods like piezo-dispensing. This facilitates time-resolved cryo-EM for studying short-lived protein states.

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Preparation of High-Temperature Sample Grids for Cryo-EM
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Preparation of High-Temperature Sample Grids for Cryo-EM

Published on: July 26, 2021

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Area of Science:

  • Structural Biology
  • Biophysics
  • Biochemistry

Background:

  • Cryo-electron microscopy (cryo-EM) is advancing with new hardware and algorithms, yielding higher resolution structures.
  • Traditional cryo-EM grid preparation methods are being replaced by faster techniques.
  • Time-resolved cryo-EM requires rapid sample mixing and freezing to capture transient states.

Purpose of the Study:

  • To detail a protocol for rapid cryo-EM grid preparation using an in-house device.
  • To enable standard fast grid preparation and time-resolved cryo-EM experiments.
  • To provide a foundation for others developing similar devices and time-resolved studies.

Main Methods:

  • Utilized an in-house time-resolved cryo-EM device for grid preparation.
  • Employed methods such as piezo-electric dispensers, pin printing, and direct spraying.
  • Developed a protocol requiring minimal sample volume (≥ 50 µL) and concentration (≥ 2 mg/mL).

Main Results:

  • Achieved grid preparation speeds from seconds down to milliseconds.
  • Enabled sample application to freezing delays as low as 10 ms for time-resolved studies.
  • Observed increased ice thickness at faster speeds compared to traditional blotting methods.

Conclusions:

  • The developed protocol significantly accelerates cryo-EM grid preparation.
  • This advancement is crucial for time-resolved cryo-EM, allowing the study of transient molecular states.
  • The protocol serves as a valuable resource for researchers designing custom grid preparation devices and time-resolved experiments.