A FRET-based near-infrared ratiometric fluorescent probe for detection of mitochondria biothiol

A new fluorescent probe A with BODIPY as FRET donor and near-infrared rhodamine as FRET acceptor is constructed through disulfide bonding and use for ratiometric fluorescence detection of biothiol. Due to the efficient fluorescence resonance energy transfer (FRET) from BODIPY donor to near-infrared rhodamine acceptor, Probe A only displays near-infrared rhodamine fluorescence (λ_em = 656 nm) under BODIPY excitation at 480 nm. The presence of biothiol leads to BODIPY fluorescence increases (λ_em = 511 nm) and near-infrared rhodamine fluorescence decreases since the disulfide bond of the probe is broken by biothiols, effectively separating the donor from the acceptor, thus inhibiting the FRET process. Probe A exhibits remarkable high selectivity and excellent linear relationship from 10 μM to 100 μM of GSH, with low detection limit as 0.26 μM. Cellular imaging experiments shows that the probe is predominantly present in mitochondria and has been successfully applied to detect biothiol concentrations changes in mitochondria of living cells.