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UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258
DNA-bound Hoechst 33258 is readily excited with UV light and emits blue fluorescence, however, upon exposure to UV, the dye undergoes photobleaching as well as photoconversion to a blue-excited green-emitting form. We demonstrate that the UV-generated green-emitting form of Hoechst 33258 exhibits sp...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4246133/ https://www.ncbi.nlm.nih.gov/pubmed/25312832 http://dx.doi.org/10.1007/s10895-014-1468-y |
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author | Żurek-Biesiada, Dominika Waligórski, Piotr Dobrucki, Jurek W. |
author_facet | Żurek-Biesiada, Dominika Waligórski, Piotr Dobrucki, Jurek W. |
author_sort | Żurek-Biesiada, Dominika |
collection | PubMed |
description | DNA-bound Hoechst 33258 is readily excited with UV light and emits blue fluorescence, however, upon exposure to UV, the dye undergoes photobleaching as well as photoconversion to a blue-excited green-emitting form. We demonstrate that the UV-generated green-emitting form of Hoechst 33258 exhibits spectral properties very similar to the form of the dye that can be obtained by subjecting it to an acidic environment (pH 0.5–3.0). We also demonstrate that exposure of Hoechst 33258 to UV light (or hydrogen peroxide) leads to generation of the protonated (1+, 2+, 3+ and possibly the 4+) forms of the dye. Photoconversion of Hoechst 33258 has recently been exploited in single molecule localisation microscopy, thus understanding photophysics of this process can facilitate further development of high resolution optical imaging. |
format | Online Article Text |
id | pubmed-4246133 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-42461332014-12-02 UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 Żurek-Biesiada, Dominika Waligórski, Piotr Dobrucki, Jurek W. J Fluoresc Original Paper DNA-bound Hoechst 33258 is readily excited with UV light and emits blue fluorescence, however, upon exposure to UV, the dye undergoes photobleaching as well as photoconversion to a blue-excited green-emitting form. We demonstrate that the UV-generated green-emitting form of Hoechst 33258 exhibits spectral properties very similar to the form of the dye that can be obtained by subjecting it to an acidic environment (pH 0.5–3.0). We also demonstrate that exposure of Hoechst 33258 to UV light (or hydrogen peroxide) leads to generation of the protonated (1+, 2+, 3+ and possibly the 4+) forms of the dye. Photoconversion of Hoechst 33258 has recently been exploited in single molecule localisation microscopy, thus understanding photophysics of this process can facilitate further development of high resolution optical imaging. Springer US 2014-10-14 2014 /pmc/articles/PMC4246133/ /pubmed/25312832 http://dx.doi.org/10.1007/s10895-014-1468-y Text en © The Author(s) 2014 https://creativecommons.org/licenses/by/4.0/ Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Paper Żurek-Biesiada, Dominika Waligórski, Piotr Dobrucki, Jurek W. UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title | UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title_full | UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title_fullStr | UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title_full_unstemmed | UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title_short | UV-induced Spectral Shift and Protonation of DNA Fluorescent Dye Hoechst 33258 |
title_sort | uv-induced spectral shift and protonation of dna fluorescent dye hoechst 33258 |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4246133/ https://www.ncbi.nlm.nih.gov/pubmed/25312832 http://dx.doi.org/10.1007/s10895-014-1468-y |
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