DNA double staining for a fluorescence energy transfer study of chromatin in liver cells.

Cell Biophysics
Giovanni BottiroliS Barni

Abstract

Methodological aspects related to the application of techniques based on fluorescence energy transfer in the study of chromatin structure, were first examined. Fluorochromes specific for DNA with different interaction mechanisms were employed, both in single and double stainings. The following dye pairs were considered as donor/acceptor couples: Hoechst 33342 or DAPI/Mithramycin A or Chromomycin A3, Hoechst 33342 or DAPI/Propidium Iodide, and Mithramycin A or Chromomycin A3/Propidium Iodide. Spectrofluorometric analysis showed that the spectral distribution of the dye pair Ho/PI is more suitable for the evaluation of energy transfer efficiency. This dye pair was used in the study of the chromatin microstructure in rat hepatocytes isolated from livers at two different growth stages. In particular, diploid mono- and binucleated cells from young and adult rats were considered. The results indicated the existence of a more homogeneous situation in young than in adult rats. In the latter case, the statistical analysis indicates the presence of two groups of energy transfer values. The different efficiency values in energy transfer can be considered a consequence of chromatin structure rearrangement and are tentatively interpreted ac...Continue Reading

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Citations

Mar 13, 1992·Journal of Photochemistry and Photobiology. B, Biology·G BottiroliR Ramponi
Jun 18, 2003·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Shin-Ichi MurataRyohei Katoh
Oct 19, 2005·Journal of Biomedical Optics·Shin-ichi MurataRyohei Katoh
Jan 1, 1992·Biophysical Chemistry·A MazziniG Sartor

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