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Journal of Histochemistry and Cytochemistry, Vol. 46, 389-396, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Fine Structural Specific Visualization of RNA on Ultrathin Sections

Marco Biggiogeraa,b and Stanislav Fakana
a Centre of Electron Microscopy, University of Lausanne, Lausanne, Switzerland, Dipartimento di Biologia Animale, Laboratorio di Istologia
b Centro di Studio per l'Istochimica del CNR, University of Pavia, Pavia, Italy

Correspondence to: Marco Biggiogera, Dipartimento di Biologia Animale, Laboratorio di Istologia, U. Of Pavia, Piazza Botta 10, 27100 Pavia, Italy.

We describe a new technique that allows specific visualization of RNA at the electron microscopic level by means of terbium citrate. Under the conditions presented here, terbium binds selectively to RNA and stains nucleoli, interchromatin granules, peri-chromatin fibrils, perichromatin granules, and coiled bodies in the cell nucleus, whereas ribosomes are the only contrasted structures in the cytoplasm. All the cell components contrasted by terbium are known to contain RNA. When ultrathin sections are pretreated with RNase A or nuclease S1 (specific for single-stranded nucleic acids), staining does not occur. Neither DNase nor pronase influences the reaction. We conclude that terbium staining is selective for RNA and especially for single-stranded RNA. The staining can be performed on thin sections of material embedded both in epoxy and in acrylic resins. The technique is not influenced by the aldehyde fixative used and can also be utilized after immunolabeling. The endproduct is very fine and, although weak in contrast, is suitable for high-resolution observations. (J Histochem Cytochem 46:389–395, 1998)

Key Words: electron microscopy, terbium, lanthanides, RNA staining, cytochemistry


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