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Related Concept Videos

Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Updated: Jul 5, 2025

Author Spotlight: Enhancing Cryo-EM Sample Preparation with Streptavidin-Biotin Approach
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Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation.

Trinity Cookis1, Paul Sauer2, Simon Poepsel3

  • 1Department of Molecular and Cell Biology, University of California, Berkeley; trinity_cookis@berkeley.edu.

Journal of Visualized Experiments : Jove
|January 15, 2024
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Summary

This study presents a protocol for fabricating streptavidin affinity grids, enhancing cryo-electron microscopy (cryo-EM) sample preparation by preventing denaturation and improving orientation. These grids simplify sample handling for the cryo-EM community.

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Optimizing Sample Preparation for Cryogenic Electron Microscopy
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Area of Science:

  • Structural Biology
  • Biophysics
  • Microscopy

Background:

  • Cryo-electron microscopy (cryo-EM) faces challenges with sample preparation, including denaturation and orientation issues at the air-water interface.
  • Current cryo-EM labs rarely use streptavidin affinity grids due to lack of commercial availability and complex fabrication.
  • These grids offer a solution by protecting samples from the air-water interface and aiding in sample concentration.

Purpose of the Study:

  • To provide a detailed, optimized protocol for fabricating streptavidin affinity grids.
  • To make streptavidin affinity grids more accessible to researchers for cryo-EM and negative-stain experiments.
  • To address common fabrication challenges through a troubleshooting guide.

Main Methods:

  • Growing two-dimensional streptavidin crystals onto a biotinylated lipid monolayer.
  • Applying the monolayer directly to standard holey-carbon cryo-EM grids.
  • Utilizing the high-affinity streptavidin-biotin interaction for sample binding.

Main Results:

  • A robust protocol for fabricating streptavidin affinity grids is established.
  • The grids facilitate the binding of biotinylated samples, protecting them from air-water interface effects.
  • The method allows for sample concentration and purification directly on the grids.

Conclusions:

  • The developed protocol enables robust fabrication of streptavidin affinity grids.
  • These grids significantly improve cryo-EM sample preparation by mitigating common issues.
  • The accessibility of streptavidin affinity grids is enhanced for the broader cryo-EM research community.