Fluorescent Dyes For Detecting Hydrogen Sulfide In Cells

As the simplest biothiol,the studies of the mechanism of hydrogen sulfide to participate in the physiological activities is still the hotspot of scientific research.But,it is insufficient and there is great demand for developing new technology to better detects hydrogen sulfide.Meanwhile,traditional analytical techniques have great application limitations in selective detection of hydrogen sulfide in living cells and tissues.In contrast,fluorescent imaging can achieve the detection of hydrogen sulfide with high sensitivity,selectivity and biological compatibility.Thus,the design of fluorescent dyes for better detects hydrogen sulfide in cells will be contribute to the better understanding of the mechanism of hydrogen sulfide.The following work were mainly carried out in this work.1.The fluorescent dye Lyso-NA was designed and synthesized based on naphthalimide,it features large Stokes shift,superior photostability,indepengent on pH and suitable for specific lysosome imaging.2.In view of its strong nucleophilicity and the selective addition towards the quaternized benzothiazole C-2 atom,a fluorescent dye BOC was designed and synthesized for ratiometric detecting of H2 S.BOC is a near-infrared fluorescent dye with large Stokes shift and molar extinction coefficient.The value of log(I492/I652)and concentrations of hydrogen sulfide represented a good linear relationship and demenstrated BOC is a ratiometric dye.BOC possessed high selectivity and anti-interference over other interfering ions and thiols,a short response time of 45 s.BOC was cell permeable and has potential for the imaging of H2 S in cells.3.Mito-BT is also a near-infrared fluorescent dye depends on triphenylamine and possessed large Stokes shift.The intensity ratio(I525/I645)increased linearly with the addition of hydrogen sulfide,which indicated Mito-BT can realize the ratiometric detection of it with a high sensitivity.Furthermore,Mito-BT behaved an intense response for hydrogen sulfide and illustrated its excellent selectivity and anti-interference.The response time of Mito-BT for hydrogen sulfide was rapid and a balance was reached after 10 s.The co-localization study showed that Mito-BT can selectively stains mitochondria and can be used for detection of hydrogen sulfide in cells.4.Based on the studies of Mito-BT,Mito-CZ was constructed for targeting mitochondria and ratiometric detection of hydrogen sulfide.The testing shows that Mito-CZ owns large Stokes shift and molar extinction coefficient and in favor of its biological imaging applications for hydrogen sulfide.The intensity ratio displayed a good increasing linear relationship toward hydrogen sulfide with high sensitivity.Besides,Mito-CZ showed good selectivity and anti-interference capability.The counter staining demonstrated Mito-CZ can selective stains mitochondria and can be used for detection of hydrogen sulfide in cells.Four fluorescent dyes Lyso-NA,BOC,Mito-BT and Mito-CZ were designed and synthesized by simple methods in this paper.The structures of these dyes were characterized by 1H NMR,13 C NMR and MS spectra,and their optical properties were investigated by UV-vis absorbance and fluorescence emission spectra.Lyso-NA can be used for specific stains lysosomes.Three other fluorescent dyes realized the ratiometric detection of hydrogen sulfide with a good performance utilize its strong nucleophilicity,all of them can be applicated as tools for detection of hydrogen sulfide in cells and has certain theoretical research and practical application values.