A two-photon fluorescent probe for biological Cu (Ⅱ) and PPi detection in aqueous solution and in vivo

The first two-photon fluorescent probe (PC) for selectively detecting biological Cu (Ⅱ) and pyrophosphate (PPi) has been developed based on 7-substituted coumarin in this study. The probe presented excellent selective two-photon "on-off-on" detection signal for Cu(II) /PPi in aqueous solution. The two-photon detection ensemble (PCCu) can detect PPi released from DNA amplification after the polymerase chain reactions (PCR). The probe showed low cytotoxicity and good biocompatibility, and therefore can be applied for imaging Cu(II)/PPi in living cells under two-photon excitation. Furthermore, the ensemble probe (PCCu) was also used to image PPi in deep living rat tissues (~100µm) and in 5-days old zebrafish.