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Related Experiment Videos

Controlled environment vitrification system: an improved sample preparation technique.

J R Bellare1, H T Davis, L E Scriven

  • 1Department of Chemical Engineering & Materials Science, University of Minnesota, Minneapolis 55455-0132.

Journal of Electron Microscopy Technique
|September 1, 1988
PubMed
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The controlled environment vitrification system (CEVS) enables cryofixation of hydrated samples, preventing artifacts from uncontrolled environments. This method preserves microstructure for accurate analysis of biological and colloidal systems.

Area of Science:

  • Cryo-electron microscopy
  • Materials science
  • Biophysics

Background:

  • Specimen preparation in uncontrolled environments can introduce artifacts due to evaporation and heat transfer.
  • Vitrification of hydrated samples at non-ambient temperatures (e.g., in vivo or elevated temperatures) is challenging.
  • Maintaining sample integrity during preparation and cryofixation is crucial for accurate microstructure analysis.

Purpose of the Study:

  • To develop and present a controlled environment vitrification system (CEVS) for preserving hydrated samples.
  • To enable cryofixation of biological and colloidal dispersions and aggregates under controlled temperature and saturation.
  • To minimize artifacts during specimen preparation and vitrification for advanced microscopy.

Main Methods:

Related Experiment Videos

  • Development of a chamber system for controlled temperature and chemical activity during specimen preparation.
  • Utilizing a plunger system for rapid specimen propulsion into a cryogenic bath.
  • Synchronous shutter mechanism for near-instantaneous vitrification upon plunging.

    • Main Results:

    • Successful cryofixation of hydrated samples, preserving original microstructure.
    • Demonstration of artifact avoidance in TEM micrographs of surfactant microstructures.
    • Validation of the CEVS for maintaining sample integrity during vitrification.

    Conclusions:

    • The CEVS effectively prevents artifacts associated with uncontrolled laboratory atmospheres.
    • This technique is valuable for studying hydrated biological and colloidal systems at various temperatures.
    • The CEVS has broad applications for SEM, freeze-fracture, and in-situ dynamic process studies.