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Related Experiment Videos

Embedment-free section electron microscopy.

Hisatake Kondo1

  • 1Division of Histology, Department of Cell Biology, Graduate School of Medicine, Tohoku University, Aobaku, Sendai, Japan. hkondo@mail.tains.tohoku.ac.jp

Journal of Electron Microscopy
|December 23, 2006
PubMed
Summary

Embedment-free electron microscopy using polyethylene glycol (PEG) allows clearer viewing of biological structures. This technique revealed microtrabecular lattices, suggesting cytoplasm behaves like a protein solution, influencing organelle movement.

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Traditional epoxy embedding hinders clear viewing of biological structures with similar electron densities.
  • Need for advanced microscopy techniques to clarify cellular structures and intracellular environments.

Purpose of the Study:

  • To introduce and validate embedment-free section electron microscopy for biological specimens.
  • To investigate the nature and behavior of intracellular microtrabecular lattices.

Main Methods:

  • Utilized water-soluble polyethylene glycol (PEG) as a transient embedding medium.
  • Employed critical point-drying after de-embedding PEG with water.
  • Examined semithin sections using electron microscopy.

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Main Results:

  • Observed microtrabecular lattices with varying compactness in different cellular domains.
  • Reproduced similar lattices in vitro from protein solutions, correlating compactness with concentration.
  • Demonstrated lattice compactness changes with osmotic shock and identified intracellular protein localization.

Conclusions:

  • Microtrabeculae represent the presence of proteins in the cytoplasm, which acts as an aqueous solution.
  • Varying lattice compactness suggests sol-to-gel state transitions within cytoplasmic domains.
  • Intracellular protein concentration and sol/gel states, alongside cytoskeleton, may control organelle localization and movement.