MRC/CORU serial number P1.083.
Article first published online: 30 MAR 2005
Copyright © 1985 Wiley-Liss, Inc.
Volume 6, Issue 4, pages 310–315, July 1985
How to Cite
Watson, J. V., Nakeff, A., Chambers, S. H. and Smith, P. J. (1985), Flow cytometric fluorescence emission spectrum analysis of hoechst-33342-stained DNA in chicken thymocytes. Cytometry, 6: 310–315. doi: 10.1002/cyto.990060406
This work was partially supported by research grants R01 AM28587 awarded to A.N. by the National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases, NIH, DHHS, and research Career Development Award K04 HL00440 awarded to A.N. by the National Heart, Lung and Blood Institute, NIH, DHHS.
- Issue published online: 16 JUN 2005
- Article first published online: 30 MAR 2005
- Manuscript Accepted: 3 FEB 1985
- Manuscript Received: 6 AUG 1984
- Hoechst 33342;
- DNA binding;
- fluorescence spectrum
Hoechst-33342-stained chicken thymocytes were analysed simultaneously on two fluorescence wavelength bands (green and violet) in our custom-built flow cytometer, and two major subsets were identified. In one subset (33% of the total) the emission spectrum remained constant with time, with little change in the respective green and violet fluorescence intensities. In the other subset (42% of the total) the green fluorescence increased during staining, resulting in a considerable change in the green-to-violet ratio, due to a change in the “shape” of the fluorescence emission with time. The data indicate that two binding sites, or two types of binding at the same site, exist in DNA for this dye and that these have different binding energies and, consequently, different fluorescence emission properties.