A dual-mode strategy for sensing and bio-imaging of endogenous alkaline phosphatase based on the combination of photoinduced electron transfer and hyperchromic effect.

Benefit from the additional correction of the output signal in dual-mode detection, traditional dual signal readout strategies are performed by constructing the ratiometric fluorescent probe through excitation energy transfer (EET) or fluorescence resonance energy transfer (FRET). To avoid the complicated modification process and obtain the results rapidly, a simple dual-mode sensing strategy based on the electronic effects of p-nitrophenol (PNP) is described to monitor the activities of alkaline phosphatase (ALP). In the sensing platform, p-nitrophenylphosphate was used as a substrate to produce the PNP using ALP as the catalyst. Due to the PNP possesses negative effect of induction and conjugation, photoinduced electron transfer and hyperchromic effect have been achieved between PNP and polyethyleneimine-protected copper nanoclusters (PEI-Cu NCs), which caused the changes of the fluorescence intensity and UV-visible absorption. The dual-mode signal sensing system showed the satisfactory linear results of ALP from 1 to 100 U/L for fluorescent sensing strategy and 1-70 U/L for the absorption method with a competitive LOD of 0.27 and 0.87 U/L (signal-to-noise ratio of 3). This strategy detected biological ALP in human serum and bio-imaging of endogenous ALP in A549 cells successfully, which verifies a certain potential of the strategy for the practical applications.