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Microwave processing for scanning electron microscopy.

F Hernández1, R Guillén

  • 1Facultad de Microbiología and Unidad de Microscopia Electrónica, Universidad de Costa Rica, San José, Costa Rica. hchavarr@cariari.ucr.ac.cr

European Journal of Morphology
|March 1, 2000
PubMed
Summary

This study introduces a rapid microwave-assisted method for Scanning Electron Microscopy sample preparation, reducing processing time from 8 hours to 1 hour. The technique effectively preserves ultrastructural details in diverse biological specimens.

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Area of Science:

  • Microscopy
  • Electron Microscopy
  • Biological Sciences

Background:

  • Standard Scanning Electron Microscopy (SEM) sample preparation is time-consuming, typically requiring at least 8 hours.
  • Conventional methods involve aldehyde fixation, osmium postfixation, ethanol dehydration, and drying.

Purpose of the Study:

  • To develop a significantly faster method for preparing biological samples for SEM.
  • To evaluate the efficacy of microwave irradiation in accelerating the SEM sample preparation process.
  • To assess the preservation of ultrastructural details using the accelerated method.

Main Methods:

  • Biological samples (mosquitoes, protozoa, bacteria, murine tissues) were processed using a domestic microwave oven.
  • Microwave treatment was applied for 2 minutes at 20% maximum power during the fixation and dehydration steps.

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  • Samples were fixed with aldehyde, postfixed with osmium, and dehydrated in graded ethanol.

    • Main Results:

    • The complete sample preparation procedure, from fixation to 100% ethanol dehydration, was reduced to approximately 1 hour.
    • Ultrastructural details of various biological specimens, including Aedes mosquitoes, Tritrichomonas muris, Clostridium oceanicum, and murine liver/intestine, were well-preserved.
    • The microwave-assisted method demonstrated comparable or superior preservation compared to conventional techniques in terms of speed.

    Conclusions:

    • Microwave-assisted sample preparation offers a substantial time reduction for SEM.
    • This accelerated method provides good preservation of ultrastructural details across diverse biological sample types.
    • The technique is a viable alternative for rapid SEM analysis, particularly in research settings requiring quick turnaround times.