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An FITC‐BODIPY FRET Couple: Application to Selective, Ratiometric Detection and Bioimaging of Cysteine
Author(s) -
Ma Dong Hee,
Kim Dokyoung,
Akisawa Takuya,
Lee KyungHa,
Kim KyongTai,
Ahn Kyo Han
Publication year - 2015
Publication title -
chemistry – an asian journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 106
eISSN - 1861-471X
pISSN - 1861-4728
DOI - 10.1002/asia.201403073
Subject(s) - förster resonance energy transfer , bodipy , cysteine , chemistry , fluorescein , fluorescence , moiety , biophysics , confocal microscopy , photochemistry , biochemistry , enzyme , stereochemistry , physics , quantum mechanics , biology , microbiology and biotechnology
A novel FRET couple of fluorescein is disclosed, and it was readily constructed by conjugating an amino‐BODIPY dye, a new FRET donor, with fluorescein isocyanate. Its potential was demonstrated by a fluorescence sensing system for cysteine, which was prepared by introducing acryloyl groups to the fluorescein moiety. The FRET probe exhibited promising ratiometric response to cysteine with high selectivity and sensitivity in a buffer solution containing acetonitrile at a physiological pH of 7.4, but showed slow reactivity. This slow response was solved by addition of a surfactant, thus allowing ratiometric imaging and determination of the endogenous level of cysteine in cells in HEPES buffer, by confocal fluorescence microscopy. Imaging experiments toward various cells suggested that such aryl acrylate type probes are vulnerable to the ubiquitous esterase activity. For the selected C6 cell line, in which the esterase activity was minimal, the ratiometric quantification of cysteine level was demonstrated. The FRET probe was also applied to determine the level of cysteine in human blood plasma.