Design And Synthesis Of Fluorescent Probe Based On Quantum Dots And Its Preliminary Application In Living Cells

There are mainly four parts in this thesis:I. Preface.Ⅱ. The synthesis and chracterization of the fluorescent probe Quinazoline-QD.Ⅲ. The synthesis and chracterization of the fluorescent probe Phenylephrine-QD.Ⅳ. Preliminary application of fluorescent probe Quinazoline-QD.In chapterⅠ, the significance, research method and development of single molecule research in single cell were introduced. Then,α1-adrenoceptor and its research development were introduced. It also disccussed the fluorescent methods of studyingα1-adrenoceptors, and make a comparison between traditional dyes, green fluorescent proteins and quantum dots. It also introduced the applications of quantum dots in biological study. Then the detection methods of fluorescence was introduced, especially the total internal reflection fluorescence microscopy.In chapterⅡ, the design and synthesis of fluorescent probe for labelingα1-adrenoceptors in living cell were described. To reduce the steric hindrance of QDs, we add a PEG chain between the small drug molecule and QDs. So it makes the movement of the drug molecule more flexible, which is closer to the movement of its own. The synthesis includes six parts:the synthesis of biotin N-hydroxysuccinimide ester, the synthesis of 2-piperazine-4-amino-6,7-dimethoxyquinazoline, the synthesis of heterobifunctional PEG (H2N-PEG-COOH), the synthesis of heterobifunctional PEG (Bio-PEG-NHS), the synthesis of biotin 2-piperazine-4-amino-6,7-dimethoxyq-uinazoline and the synthesis of fluorescent probe Quinazoline-QD. The final product was chracterized by 1H NMR and MALDI-TOF mass spectrometry.In chapterⅢ, we used phenylephrine, an agonist ofα1-adrenoceptors to decorate the QDs. Due to its active anmino group and the different group which PEG was linked, the synthetic route employed was different from 2-piperazine-4-amino-6, 7-dimethoxyquinazoline. At first, a protective group di-tert-butyl dicarbonate was used. It is very effective in protecting amino group. It was removed by trifluoroacetic acid, which remained CO2 as a byproduct and had no side effect for the product. The final product was chracterized by 1H NMR and MALDI-TOF mass spectrometry. In the last part, we accessed the specific interaction between Quinazoline-QD andα1-adrenoceptors in living cells via total internal reflection fluorescence microscopy as a preliminary application. The results showed that there were a lot of fluorescent dots binding to the membranes of cells which expressedα1B-adrenoceptors, and there were little fluorescent dots on the surface of cells which did not expressα1B-adrenoceptors. It demonstrated that the probe could specifically bind toα1-adrenoceptors and proved the distribution of this kind ofα1-adrenoceptors.