| Comparing with the conventional techniques including colorimetric method,electrochemical analysis and chromatographic analysis,fluorescent techniques,such as fluorescent protein labeling,fluorescent probe detection,quantum dot analysis,are more widely used in the bio-analysis,owing to their advantages in terms of free-sample pretreatment,visualization research,non-invasive detection and so on.Usually,the fluorescent probe can be designed by combining a fluorophore with a recognition group.In the bio-system,there are many important signaling molecules,among which the endogenous hydrogen sulfide plays the vital role in the life course.At normal physiological level,hydrogen sulfide(H2S)can participate in the regulation of many life activities,including neuromodulation,angiogenesis,cell metabolism and insulin secretion,etc.In addition to hydrogen sulfide,the intracellular small molecule mercapto compounds(bio-mercaptan,-SH)also play an extremely important role in life activities.In the human body,content of bio-mercaptan and the related balance directly determine that whether the whole body mechanism is healthy.Besides,there exist a number of polysulfides(-S-S-)in the living body.It is reported that trace cyanide does not cause human poisoning.The reason lies in that the polysulfides in body can act as an antidote for cyanide.Therefore,the polysulfide plays an important role in the physiological activities of living organisms.As the above three types of important molecules(H2S,-SH and-S-S-)play the important role in the life activities,it is significant for researchers to design the simple,convenient,rapid and selective fluorescent probe for the detection of them in cells or in vivo.In this dissertation,we designed three near infrared(NIR)probes for the detection of H2 S,-SH and-S-S-compounds and applied the developed probes to the fluorescence imaging of targets in living cells.The main content is as follows:Chapter 1: The origin,development,luminescence principle and the recognition mechanism are introduced.The research status of the probe is summarized.The background of mercapto compounds was reviewed,and some ideas and plans were proposed.Chapter 2: The synthesis and application of fluorescent probe?(DCYMB)(E)-2-(2-(4-(2,4-dinitrophenoxy)styryl)-4H-chromen-4-ylidene)malononitrile.DCYMB was synthesized based on the reaction of dinitrobenzene methyl-benzopyran(DCMB)(fluorophore)and2,4-dinitrofluorobenzene(reporting group).The developed probe itself does not release the fluorescence.Upon addition of hydrogen sulfide(H2S),the fluorescence emission(680 nm)was observed.The probe DCYMB proves to be sensitive(28 nM),selective and low toxic(85%).With DCYMB,the fluorescence imaging of H2 S in living HeLa cells was successfully carried out.Chapter 3: The synthesis and application of fluorescent probe?(SDID)3'-oxo-3'H-spiro[dibenzo[c,h]xanthene-7,1'-isobenzofuran]-3,11-diylbis(3,5-dinitrobenzenesulfonate).SDID was synthesized by combing the naphthol fluorescein(as fluorophore)and2,4-dinitrobenzenesulfonyl(as mercapto-recognition group).The developed probe itself does not release the fluorescence.Upon addition of-SH,the fluorescence emission(640 nm)was observed.The probe SDID proves to be sensitive(15 nM),selective and low toxic(85%).With SDID,the fluorescence imaging of-SH compounds in living HeLa cells was successfully carried out.Chapter 4: The synthesis and application of fluorescent probe ?(DCPMB)(E)-4-(2-(4-(dicyanomethylene)-4H-chromen-2-yl)vinyl)phenyl 2-hydroselenobenzoate ? DCPMB was synthesized using the dinitrobenzene methyl-benzopyran(DCMB)(as fluorophore)and the2,2'-di-selenium dibenzoic acid(as reporting group).The developed probe itself does not release the fluorescence.Upon addition of –S-S-compounds,the fluorescence emission(663 nm)was observed.The probe DCPMB proves to be sensitive(63 nM),selective and low toxic(90%).With DCPMB,the fluorescence imaging of-SH compounds in living HeLa cells was successfully carried out.Chapter 5: Conclusions. |
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