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Tuned lifetime, at the ensemble and single molecule level, of a xanthenic fluorescent dye by means of a buffer-mediated excited-state proton exchange reaction

Tuned lifetime, at the ensemble and single molecule level, of a xanthenic fluorescent dye by means of a buffer-mediated excited-state proton exchange reaction
Tuned lifetime, at the ensemble and single molecule level, of a xanthenic fluorescent dye by means of a buffer-mediated excited-state proton exchange reaction
The photophysical behaviour of the new fluorescein derivative 9-[1-(2-methyl-4-methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one has been explored by using absorption, and steady-state, time-resolved and single-molecule fluorescence measurements. The apparent ground-state acidity constant of the dye determined by both the absorbance and steady-state fluorescence is almost independent of the added buffer and salt concentrations. The excited-state proton exchange reaction around the physiological pH becomes reversible upon addition of phosphate buffer, inducing a pH-dependent change of the steady-state fluorescence and decay times. Fluorescence decay traces, collected as a function of total buffer concentration and pH, were analyzed by global compartmental analysis (GCA) to elucidate the values of the excited-state rate constants. The features of this system make the fluorescence decays monoexponential at pH values and phosphate buffer concentrations higher than 6.10 and 0.2 M respectively, with the possibility of tuning the fluorescence lifetime value by changing pH or buffer concentrations. The tuned lifetimes obtained by means of phosphate concentration at constant pH have also been recovered at the single-molecule level.
1463-9076
5400-5407
Paredes, Jose M.
56f39307-ce21-4a3c-9ec5-61dfc850ab43
Crovetto, Luis
45d4cd97-3fe5-4498-a861-bc06492eb09d
Rios, Ramon
609bedf2-e886-4d62-a676-a32b6f8c1441
Orte, Angel
d841ace2-236b-4fcd-b29d-5cf59ebe5408
Alvarez-Pez, Jose M.
6970f24c-ecb5-418b-9847-f7a6be1d91f2
Talavera, Eva M.
e0319787-e90f-48dd-b89f-5c631bbe15a1
Paredes, Jose M.
56f39307-ce21-4a3c-9ec5-61dfc850ab43
Crovetto, Luis
45d4cd97-3fe5-4498-a861-bc06492eb09d
Rios, Ramon
609bedf2-e886-4d62-a676-a32b6f8c1441
Orte, Angel
d841ace2-236b-4fcd-b29d-5cf59ebe5408
Alvarez-Pez, Jose M.
6970f24c-ecb5-418b-9847-f7a6be1d91f2
Talavera, Eva M.
e0319787-e90f-48dd-b89f-5c631bbe15a1

Paredes, Jose M., Crovetto, Luis, Rios, Ramon, Orte, Angel, Alvarez-Pez, Jose M. and Talavera, Eva M. (2009) Tuned lifetime, at the ensemble and single molecule level, of a xanthenic fluorescent dye by means of a buffer-mediated excited-state proton exchange reaction. Physical Chemistry Chemical Physics, 11 (26), 5400-5407. (doi:10.1039/b820742a). (PMID:19551208)

Record type: Article

Abstract

The photophysical behaviour of the new fluorescein derivative 9-[1-(2-methyl-4-methoxyphenyl)]-6-hydroxy-3H-xanthen-3-one has been explored by using absorption, and steady-state, time-resolved and single-molecule fluorescence measurements. The apparent ground-state acidity constant of the dye determined by both the absorbance and steady-state fluorescence is almost independent of the added buffer and salt concentrations. The excited-state proton exchange reaction around the physiological pH becomes reversible upon addition of phosphate buffer, inducing a pH-dependent change of the steady-state fluorescence and decay times. Fluorescence decay traces, collected as a function of total buffer concentration and pH, were analyzed by global compartmental analysis (GCA) to elucidate the values of the excited-state rate constants. The features of this system make the fluorescence decays monoexponential at pH values and phosphate buffer concentrations higher than 6.10 and 0.2 M respectively, with the possibility of tuning the fluorescence lifetime value by changing pH or buffer concentrations. The tuned lifetimes obtained by means of phosphate concentration at constant pH have also been recovered at the single-molecule level.

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e-pub ahead of print date: 23 April 2009
Published date: 2009
Organisations: Chemistry

Identifiers

Local EPrints ID: 347744
URI: http://eprints.soton.ac.uk/id/eprint/347744
ISSN: 1463-9076
PURE UUID: 6b851090-238e-4bf8-a0c3-c4e340f6bb4e
ORCID for Ramon Rios: ORCID iD orcid.org/0000-0002-3843-8521

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Date deposited: 29 Jan 2013 14:18
Last modified: 26 Nov 2019 01:37

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