A colorimetric and ratiometric fluorescent probe for highly selective detection of glutathione in the mitochondria of living cells

A FRET‐Based Near‐Infrared Fluorescent Probe for Ratiometric Detection of Cysteine in Mitochondria

“…Cell culture and MTT cytotoxicity assay: HeLa cells were cultured in Dulbecco'sm odified Eagle's medium (DMEM, Gibco) containing 10 %f etal bovine serum (FBS, Fisher Scientific) at 37 8Ci nahumid atmosphere containing 5% CO 2 .H eLa cells were subcultured at 80 %c onfluence using 0.25 %t rypsin (w/v) (Fisher Scientific) every other day.Astandard MTT assay was applied to determine the cytotoxicity of probe A.I nd etail, the cells were seeded in 96-well plates at an initial density of 4000 cells per well, with 100 mL DMEM medium per well. After seeding for 24 hi nt he 96-well plate, the medium was replaced by different concentrations of probe A (0, 5,10,15,25,50 mm solutions in fresh culture medium, 100 mL/well) for 48 h. After that, the cells were incubated for 4h with the tetrazolium salt dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide at af inal concentration of 500 mgmL À1 , whereupon metabolically active cells reduced the dye to the water-insoluble purple formazan dye. The dark purple crystals were dissolved with DMSO and the cell viability rate determined by measuring the absorbance at 490 nm (BioTek ELx800).…”

“…[3a, 4] However,r atiometric fluorescent probesf or cysteine detection with near-infrared emissions are currentlyl imited to af ew examples. [5] Herein, we detail an earinfrared fluorescent probe based on the FRET strategy for the ratiometric selective detection of Cys and Hcy over GSH in live cells by employing coumarin as ad onor and near-infrared rhodamine as an acceptor with ap iperazine-tethered spacer. We chose coumarin and the near-infrared rhodamine derivative because of their excellent optical properties, including outstanding photostability,h igh absorption coefficients, and high fluorescenceq uantumy ields.…”

Nile-Red-Based Fluorescence Probe for Selective Detection of Biothiols, Computational Study, and Application in Cell Imaging

“…Colorimetric and fluorescent probes have become a new hotspot owing to their remarkable advantages in fundamental study of analytical chemistry and photochemistry [ 29 , 30 , 31 , 32 , 33 , 34 , 35 ]. To date, a lot of colorimetric and fluorescent probes for biothiols have been developed [ 36 , 37 , 38 , 39 , 40 ]. Latterly, scientists have begun to design new fluorescent probes based on Nile-red fluorophore for detecting biothiols, and several good probes have been reported [ 41 , 42 , 43 , 44 ].…”

Characterization of a novel styrylbenzimidazolium‐based dye and its application in the detection of biothiols

“…[ 25–27 ] In the past decades, a wide range of chemical fluorescent probes for detection of biothiols has been developed based on various mechanisms including cyclization with aldehyde, [ 28,29 ] Michael addition, [ 30–33 ] cleavage reactions by thiols, [ 34–36 ] and other reactions. [ 37–41 ] Michael addition is one of the most commonly used reactions for detection of thiol‐containing amino acids. Among the various Michael’s acceptors, maleimide moieties are in common use.…”