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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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Studying crystallisation processes using electron microscopy: The importance of sample preparation.

Martha Ilett1, Maryam Afzali1, Bilal Abdulkarim1

  • 1School of Chemical and Process Engineering, University of Leeds, Leeds, UK.

Journal of Microscopy
|April 10, 2024
PubMed
Summary
This summary is machine-generated.

Electron microscopy sample preparation methods significantly impact crystallisation studies. Cryogenic (cryo)-quenching and cryo-transmission electron microscopy (cryo-TEM) are crucial for accurate observation of early crystallisation stages.

Keywords:
crystallisationelectron microscopysample preparation

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Area of Science:

  • Materials Science
  • Crystallography
  • Electron Microscopy

Background:

  • Studying crystallisation processes requires accurate visualization of early stages.
  • Common electron microscopy sample preparation methods may introduce artefacts.
  • Understanding crystallisation kinetics and polymorph selection is vital in materials science.

Purpose of the Study:

  • To compare common electron microscopy sample preparation techniques for studying solution crystallisation.
  • To evaluate the impact of different preparation methods on crystallisation kinetics and polymorph selection.
  • To identify the most reliable method for observing early crystallisation events.

Main Methods:

  • Comparison of scanning electron microscopy (SEM) and transmission electron microscopy (TEM) sample preparation methods.
  • Investigation of calcium sulphate (gypsum) and calcium carbonate crystallisation.
  • Application of cryogenic (cryo)-quenching and full cryo-TEM techniques.

Main Results:

  • Significant differences in crystallisation kinetics and polymorph selection were observed across preparation methods.
  • Drying and chemical quenching methods were found to induce severe artefacts.
  • Artefacts can mask the true native state of crystallising solutions.

Conclusions:

  • Cryogenic (cryo)-quenching is essential for preserving the native state of crystallising solutions.
  • Full cryo-transmission electron microscopy (cryo-TEM) is the most reliable method for studying early crystallisation stages.
  • Careful selection of sample preparation is critical for accurate crystallisation research.