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FRET-based dual channel fluorescent probe for detecting endogenous/exogenous H2O2/H2S formation through multicolor images

Velusamy, Nithya, Thirumalaivasan, Natesan, Bobba, Kondapa Naidu, Podder, Arup, Wu, Shu-Pao, Bhuniya, Sankarprasad
Journal of photochemistry and photobiology 2019 v.191 pp. 99-106
cell viability, coumarin, emissions factor, fluorescence, fluorescent dyes, hydrogen peroxide, hydrogen sulfide, image analysis, neoplasm cells, neoplasms, pH, ultraviolet-visible spectroscopy, viability assays
We have developed a FRET-based fluorescent probe (PHS1) as a combination of two different fluorophores (coumarin and naphthalimide); which can detect both exogenous and endogenous H2S and H2O2 in live cells through multicolor images. The precise overlap between UV-absorption of naphthalimide and the emission band of coumarin in probe PHS1 allows the acquisition of the self-calibrated information of dual analytes through FRET-based imaging. The UV–Vis absorption (λabs 390 nm) and fluorescence emission (λem 460 nm) of probe PHS1 in the presence of H2O2 are increased ∽35- fold and ∽15-fold respectively. It also allows the estimation of the levels of H2S through enhancement of emission intensity at 550 nm. The probe PHS1 exhibits high stability against various analytes, including various pH (4–9.5). The cell viability assay data indicate that the probe is not harmful to the cancer cells. The nontoxic nature of the probe PHS1 encourages application for cancer cell labeling. The probe PHS1 can detect the level of endogenous H2O2, H2S, and H2O2/H2S in cancer cells through blue, green and FRET-based green channel imaging. PHS1 is a unique probe, has potential application for diagnosing cancer by providing information on the level of dual analytes (H2S, H2O2) in cancer cells.