Lysosome as the typical acidic organelle involved in many biological processes, the dysfunction of its pH would alter many diseases. Small-molecule fluorescent probe has been considered as vital tool for monitoring pH fluctuation in living cells. Herein, a hemicyanine based ratiometric fluorescent probe was synthesized, namely, 2,3-trimethy-3-[2-(dimethyl-amino-4-yl) vinyl]-3H-benzo[e]indole (BiDL), for rapidly detection pH under acidic conditions. BiDL exhibited ratio fluorescence emission (F534nm/F622nm) characteristic with pKa 4.25. BiDL showed good linearly response in pH 3.4-4.82, indicating that the probe can be used for quantitative detection pH. The probe also displayed large Stokes shift (112 nm/201 nm) under neutral and acidic conditions, respectively, which could effectively reduce the excitation interference. BiDL had excellent cell membrane permeability, good photo-stability and low toxicity in living cells. The dual-channel confocal fluorescent microscopic ratiometric imaging application of pH in HeLa cells and lysosome were achieved successfully, indicating that BiDL has good potential for investigating lysosome-relevant pathological and physiological processes.
Keywords: Dual-channel confocal; Fluorescent probe; HeLa cells; Large stokes shift; Lysosome; Ratiometric.
Copyright © 2019 Elsevier B.V. All rights reserved.