Abstract
Although the EDTA regressive technique allows the visualization of RNPs, this widely used method is not intended to be specific for RNA alone. A fine ultrastructural visualization of RNA on ultrathin sections can be obtained with terbium citrate: this method gives a weak contrast but a very fine end product which allows observations at a high resolution level.
The procedure is very simple since it consists only of a period of incubation and very short washes, which are the crucial point of this technique to avoid the Tb removal from RNA. This method does not require any special type of fixation and embedding.
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References
Bernhard M (1969) A new staining procedure for electron microscopical cytology. J Ultrastruct Res 27:250–265
Chan-Curtis V, Belt VD, Ladoulis CT (1970) Cytochemical localization of nucleic acids by acriflavine-phosphotungstate complex for fluorescence microscopy and electron microscopy. J Histochem Cytochem 18:609–627
Biggiogera M, Flach Biggiogera F (1989) Ethidium bromide- and propidium iodine-PTA staining of nucleic acids at electron microscopic level. J Histochem Cytochem 37:1161–1166
Derenzini M, Farabegoli F (1990) Selective staining of nucleic acids by osmium ammine complex in thin sections from Lowicryl-embedded samples. J Histochem Cytochem 28:1495–1501
Bendayan M (1982) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes: influence of fixation and embedding. Bio Cell 43:153–156
Scott JE (1973) Affinity, competition and specific interactions in the biochemistry and histochemistry of polyelectrolytes. Biochem Soc Trans 1:787–806
Ringer DP, Burchett S, Kizer DP (1978) Use of terbium fluorescence enhancement to selectively monitor DNA and RNA guanine residues and their alteration by chemical modification. Biochemistry 17:4818–4825
Hörer OL, Zaharia CN, Marcu A (1977) Terbium fluorescence in aqueous solutions of nucleic acids. Rev Roum Biochim 14:175–179
Biggiogera M, Fakan S (1998) Fine structural specific visualization of RNA on ultrathin sections. J Histochem Cytochem 46:389–395
Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–212
Ringer DP, Howell BA, Kizer DE (1980) Use of terbium fluorescence enhancement as a new probe for assessing the single-strand content of DNA. Anal Biochem 103:337–342
Acknowledgments
The authors are grateful to Ms. Francine Flach and Prof. Stan Fakan. Part of this work has been carried out at the CME, University of Lausanne, Switzerland.
This chapter is dedicated to the memory of Vàclav Kopečný and Jean-Luc Courtens.
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Biggiogera, M., Masiello, I. (2017). Visualizing RNA at Electron Microscopy by Terbium Citrate. In: Pellicciari, C., Biggiogera, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 1560. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6788-9_21
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DOI: https://doi.org/10.1007/978-1-4939-6788-9_21
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