Research On Two-photon Fluorescent Dyes For Biological Identification And Medical Diagnosis

Biomarkers are biochemical indexes, which can label the structure or functional change of organism, organs, tissues, cells and organelles. It is often used in biological identification, medical diagnosis and disease treatment. Therefore, to establish the efficient, accurate and concise method for the specific recognition of biomarkers will play an immeasurable role in biological identification, medical diagnosis and disease treatment. In recent years, fluorescence microscopic imaging and detecting technologys using fluorescent dyes for the specific recognition of biomarkers have gradually been applied to the biological identification, medical diagnosis and disease treatment. But there are still many problems in the existing fluorescent dyes for biomarkers, for example, specificity, sensitivity, biological adaptability and so on.Two kinds of targeting chloride pumps and lysosome two-photon fluorescence dyes, TPFL-ER and TPFL-Lyso, are designed and synthesized from fluorene derivatives by end-functionalization. They can locate in endoplasmic reticulum and lysosomes because of the suitable water-oil amphipathy. TPFL-ER and TPFL-Lyso could not only be used for the specific and simultaneous bioimaging of endoplasmic reticulum and lysosomes, but also decreased the interference of dye molecules to the protein movement in cells, improve the biological adaptability of the dyes.Two kinds of two-photon fluorescence dyes, sulfonylurea derivative (SU-1and SU-2), are designed and synthesized by using sulphonylurea receptor I as biomarker. SU-1and SU-2could specificity combine with sulphonylurea receptor I, which make that the fluorescence dyes have specific recognition ability for the sulphonylurea receptor I. The fluorescence dyes could successfully visualize the structural changes of endoplasmic reticulum in the process of diabetes.One kind of two-photon fluorescence dye, nitrobenzimidazole derivative (BIQ), is designed and synthesized by using specific basic group of RNA (uracil) as biomarkers. Fluorescence dye, BIQ, combine with RNA by a specific reaction between the BIQ and uracil, which make that there is an obvious "OFF-ON" fluorescence process. BIQ can specifically label RNA in vivo and in vitro, which achieves in situ real-time microscopic imaging of RNA in vivo. And more importantly, this novel RNA specificity recognition mechanism was proposed and verified by experiment. Three kinds of two-photon fluorescence dyes, indomethacin derivatives (ANQ-IMC1, ANQ-IMC2and ANQ-IMC6), are designed and synthesized by using cyclooxygenase-2(COX-2) as disease biomaker. ANQ-IMC2as contrastive molecule, Fluorescence dye, ANQ-IMC1and ANQ-IMC6can specifically combine with COX-2, which make that there is an obvious "OFF-ON" fluorescence process in vivo and in vitro. ANQ-IMC1has different wavelength fluorescence response for different concentration of COX-2, which make that ANQ-IMC1could detect and image of COX-2in ratio mode in vivo and in vitro. And the cancer tissues could be identified from nomal tissues and inflamed tissues with the naked eye after spraying ANQ-IMC1. Furthermore, the Golgi apparatus of cancer cells can be visualized by ANQ-IMC6, because that ANQ-IMC6has the suitable water-oil amphipathy.