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FRET-based dual channel fluorescent probe for detecting endogenous/exogenous H2O2/H2S formation through multicolor images
摘要： 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.
关键词： FRET-based fluorescent probe,Naphthalimide,Endogenous H2O2,Endogenous H2S,Coumarin
更新于2025-11-19 16:46:39
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Visualization of Endogenous Hydrogen Sulfide in Living Cells based on Au Nanorods@Silica Enhanced Fluorescence
摘要： Hydrogen sulfide as a gas indicator molecule plays an important role in various human physiological processes. However, due to the high volatility and diffusivity of H2S in biological systems, it is very difficult to implement a precise assay for H2S detection. Compared with the destructive instrumental methods, assays based on fluorescence probes provide noninvasive and real-time detections of H2S in living cells. In this work, we presented a fluorescent nanoprobe based on dye-functionalized Au nanorods (NRs)@silica for sensitive and selective detection of H2S in vitro and living cells. With the metal enhanced fluorescence effect, the fluorescence turn-on and turn-off were controlled by the formation and disassembly of coordination compound between dyes and copper ions. Silica matrix was used to coat the Au NRs to prevent them from the biological cytotoxicity. The effects of the different distances between Au NRs and fluorophores on fluorescent enhancement were explored and approximately 5-fold fluorescence enhancement was obtained with a distance of 22 nm. A detection of limit of 17 nM was achieved. In addition, visualization of exogenous and endogenous H2S in living cells was validated.
关键词： nanoprobes,Au nanorods@silica,metal enhanced fluorescence,living cells,endogenous H2S
更新于2025-09-04 15:30:14