Carboxy SNARF-4F as a fluorescent pH probe for ensemble and fluorescence correlation spectroscopies.
The optical spectroscopic properties of 1,4(and 5)-benzenedicarboxylic acid, 2-[10-(dimethylamino)-4-fluoro-3-oxo-3H-benzo[c]xanthen-7-yl] (carboxy SNARF-4F), a commercial promising fluorescent pH probe, are investigated in buffered aqueous solutions in the 5.6-8.2 pH range by steady-state and time-resolved spectroscopy. A multiexponential global analysis of the picosecond time-resolved data is performed. The nonprotonated A species decays monoexponentially (0.73 ns), while the protonated species AH decays following a biexponential law with time constants of 0.40 and 1.87 ns. A kinetic scheme is proposed to explain the observations, which involves AH in equilibrium with a species denoted Y. The nature of Y is discussed in terms of a possible structural change in the molecule producing the lactone form, although the formation of a hydrogen-bonded complex to the solvent cannot be ruled out. Finally, the ability of the dye to probe pH at the single-molecule level is explored using fluorescence correlation spectroscopy.
Marcotte
Nathalie
N
Van't Hoff Institute for Molecular Sciences, University of Amsterdam, Nieuwe Achtergracht 129, 1018 WS Amsterdam, The Netherlands.
Brouwer
Albert M
AM
D000042:Absorption; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008744:Methylamines / Q000737:chemistry*; D008956:Models, Chemical; D013050:Spectrometry, Fluorescence; D014966:Xanthenes / Q000737:chemistry*
C521638:1,4,-benzenedicarboxylic acid, 2-(10-(dimethylamino)-4-fluoro-3-oxo-3H-benzo(c)xanthen-7-yl); D005456:Fluorescent Dyes; D008744:Methylamines; D014966:Xanthenes