How to stain nucleic acids and proteins in Miller spreads

Lorena  Zannino; Marco Biggiogera

doi:10.4081/ejh.2022.3364

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Lorena Zannino

lorena.zannino@unipv.it

https://orcid.org/0000-0001-8889-1781

Laboratory of Cell Biology and Neurobiology, Department of Biology and Biotechnology, University of Pavia, Italy.

Marco Biggiogera

https://orcid.org/0000-0003-3834-6712

Laboratory of Cell Biology and Neurobiology, Department of Biology and Biotechnology, University of Pavia, Italy.

The spread technique proposed by Miller and Beatty in 1969 allowed for the first time the visualization at transmission electron microscopy of nucleic acids and chromatin in an isolated and distended conformation. The final step of staining the spread chromatin is of critical importance because it can strongly influence the interpretation of the results. We evaluated different staining techniques and the most part of them provided a good result. Specifically, well contrasted micrographs were obtained when staining with H₃PW₁₂O₄₀ (PTA), as originally proposed by Miller and Beatty, and with two alternatives proposed here: uranyl acetate or terbium citrate staining. Quite good contrast of the spread DNA could be achieved also by using Osmium Ammine; while no or few contrast of nucleic acids was observed by staining with KMnO₄ and H₃PMo₁₂O₄₀ (PMA) respectively.

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How to Cite

Zannino, L. ., & Biggiogera, M. (2022). How to stain nucleic acids and proteins in Miller spreads. European Journal of Histochemistry, 66(1). https://doi.org/10.4081/ejh.2022.3364

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