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Comparison of different drying procedures for scanning electron microscopy using human leukocytes

H P Ting-Beall1, D V Zhelev, R M Hochmuth

  • 1Department of Mechanical Engineering and Materials Science, Duke University, Durham, North Carolina 27708-0300, USA.

Microscopy Research and Technique
|November 1, 1995
PubMed
Summary

Comparing three drying methods for scanning electron microscopy, researchers found that air-drying with tetramethylsilane (TMS) offers the best balance of simplicity, convenience, and time savings for preparing animal cells, with no compromise on surface morphology preservation.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biotechnology

Background:

  • Scanning electron microscopy (SEM) requires proper sample preparation, including dehydration and drying, to preserve cellular structures.
  • Traditional methods like critical-point drying (CPD) are effective but can be time-consuming and complex.
  • Alternative drying agents and methods are continuously explored to optimize SEM sample preparation.

Purpose of the Study:

  • To compare the efficacy of three different drying procedures for scanning electron microscopy (SEM).
  • To evaluate surface morphology preservation and volume shrinkage in human leukocytes prepared using critical-point drying (CPD), Peldri II, and tetramethylsilane (TMS).
  • To determine the most practical and efficient method for preparing animal cells for SEM.

Main Methods:

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  • Human leukocytes were used as test specimens.
  • Three drying procedures were compared: critical-point drying (CPD), Peldri II, and air-drying with tetramethylsilane (TMS).
  • Surface morphology and volume shrinkage were assessed after each drying procedure.

Main Results:

  • All three drying procedures (CPD, Peldri II, TMS) resulted in identical preservation of surface morphology.
  • No significant difference in volume shrinkage was observed across the different dehydrants and drying techniques.
  • Air-drying with TMS demonstrated comparable results to CPD and Peldri II in terms of morphological preservation.

Conclusions:

  • Air-drying with tetramethylsilane (TMS) is a highly effective and practical method for preparing animal cells for scanning electron microscopy.
  • The TMS method offers significant advantages in simplicity, convenience, and time efficiency compared to CPD and Peldri II.
  • For routine SEM preparation of animal cells, air-drying with TMS is recommended for optimal results and workflow efficiency.