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  • Dual-fluorescence pH probe ...
    Richter, C; Schneider, C; Quick, M T; Volz, P; Mahrwald, R; Hughes, J; Dick, B; Alexiev, U; Ernsting, N P

    Physical chemistry chemical physics : PCCP, 2015-Nov-11, Letnik: 17, Številka: 45
    Journal Article

    Although seminaphtorhodafluor (SNARF) dyes are already widely used to measure pH in cells and at biofilms, their synthesis has low yield and results in an unspecific position of a carboxy-group. The separation of 5'- and 6'-carboxy-SNARF reveals a pKa difference of 0.15, calling into question pH measurements with the (commercially available) mixture. Here we replace the bulky external dicarboxyphenyl ring with a propionate group and evaluate the spectral properties of the new derivative. Proceeding to the ethyl-iodoacetamide, covalent linkage to cysteine protein sites is achieved efficiently as shown with a cyanobacterial phytochrome, extending the scarce application of SNARF in bio-labelling in the current literature. Application in fluorescence lifetime imaging is demonstrated both with the lifetime-based and ratiometric-yield method.