Cell Biophysics

, Volume 15, Issue 3, pp 249–263 | Cite as

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

  • Giovanni Bottiroli
  • Anna Cleta Croce
  • Giuseppe Gerzeli
  • Sergio Barni


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 rearrangements and are tentatively interpreted according to the functional role of the diploid cells in the two stages of liver growth.

Index Entries

Fluorescence energy transfer microspectrofluorometry DNA-fluorochromization chromatin hepatocytes liver polyploidization and binucleation 



Hoechst 33342



Mt A

Mithramycin A

Chr A3

Chromomycin A3


Propidium Iodide


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Copyright information

© Humana Press Inc. 1989

Authors and Affiliations

  • Giovanni Bottiroli
    • 1
  • Anna Cleta Croce
    • 1
  • Giuseppe Gerzeli
    • 2
  • Sergio Barni
    • 3
  1. 1.Centro di Studio per l’lstochimicaCNRItaly
  2. 2.Dipartimento di BiologiaAnimale dell’Università di PaviaItaly
  3. 3.Istituto di Fisiologia Generale e ChimicaBiologica dell’Università di SassariItaly

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