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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...

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Autores principales: Żurek-Biesiada, Dominika, Waligórski, Piotr, Dobrucki, Jurek W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2014
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.
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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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