Study of propidium iodide binding to DNA in intact cells by flow cytometry

A. Bertuzzi, I. D'Agnano, A. Gandolfi, A. Graziano, G. Starace, P. Ubezio

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Abstract

We studied the in situ binding of propidium iodide to DNA in fixed human lymphocytes, using flow cytometry. Experimental data of fluorescence emission vs dye concentration and vs cell concentration were obtained. Data were interpreted by means of two different mathematical models specific for the staining reaction, and the binding parameters were obtained by "best-fitting" of the data. A model based on two classes of binding sites with different affinity constants gave the most satisfactory fitting. The accessibility of the in situ chromatin turned out to be reduced with respect to the non in situ accessibility for ethidium bromide as reported in the literature. The present study shows the usefulness of the flow-cytometric technique for probing DNA structure in intact cells.

Original languageEnglish
Pages (from-to)257-267
Number of pages11
JournalCell Biophysics
Volume17
Issue number3
DOIs
Publication statusPublished - Dec 1990

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Keywords

  • binding models
  • chromatin
  • DNA
  • flow cytometry
  • propidium iodide

ASJC Scopus subject areas

  • Cell Biology
  • Biophysics

Cite this

Bertuzzi, A., D'Agnano, I., Gandolfi, A., Graziano, A., Starace, G., & Ubezio, P. (1990). Study of propidium iodide binding to DNA in intact cells by flow cytometry. Cell Biophysics, 17(3), 257-267. https://doi.org/10.1007/BF02990721