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Visualizing RNA at Electron Microscopy by Terbium Citrate.

Marco Biggiogera1, Irene Masiello2

  • 1Department of Biology and Biotechnology, Laboratory of Cell Biology and Neurobiology, University of Pavia, Via Ferrata 9, 27100, Pavia, Italy. marco.biggiogera@unipv.it.

Methods in Molecular Biology (Clifton, N.J.)
|February 4, 2017
PubMed
Summary

Terbium citrate provides a simple, high-resolution method for visualizing RNA ultrastructure. This technique offers fine detail for studying ribonucleoprotein complexes without specific fixation requirements.

Keywords:
CytochemistryEDTA regressive stainElectron microscopyLanthanidesRNA stainingTerbium citrate

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Microscopy

Background:

  • The EDTA regressive technique visualizes ribonucleoprotein complexes (RNPs) but lacks specificity for RNA.
  • Accurate ultrastructural visualization of RNA is crucial for understanding cellular processes.

Purpose of the Study:

  • To introduce and validate terbium citrate as a high-resolution staining method for RNA on ultrathin sections.
  • To provide a simple and effective alternative to existing RNA visualization techniques.

Main Methods:

  • Incubation of ultrathin sections with terbium citrate.
  • Short washes to prevent terbium ion (Tb) removal from RNA.
  • High-resolution electron microscopy for ultrastructural analysis.

Main Results:

  • Terbium citrate staining yields a very fine end product suitable for high-resolution imaging.
  • The method provides weak contrast but exceptional fine detail for RNA visualization.
  • The procedure is simple, requiring only incubation and brief washes, and is compatible with standard fixation and embedding.

Conclusions:

  • Terbium citrate is a valuable technique for fine ultrastructural visualization of RNA.
  • Its simplicity and high resolution make it advantageous for studying RNA localization and structure.
  • This method enhances the ability to observe RNA within cellular contexts at the ultrastructural level.