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Live Cell Imaging and Cytotoxicity of Tetraphenylethene with Aggregation-induced Emission Characteristic

Author: ZhangLei
Tutor: ZhengQiChang
School: Huazhong University of Science and Technology
Course: Hepatobiliary Surgery
Keywords: aggregation-induced emission tetraphenylethene cell imaging cytotoxicity
CLC: R318.0
Type: Master's thesis
Year: 2011
Downloads: 130
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Abstract


Background Fluorescent molecular imaging is using fluorescent molecular marker specific molecular or probe cells, research living state molecules from organization level, cell level or subcellular level and research its biological behaviour in the image aspects by qualitative and quantitative analysis. Fluorescent molecular probe imaging is widely used in biology and medical research for its high sensitivity, fast and convenient, high timeresolution and low costs. However, it is offen occurred a fluorescence abate or quenches phenomenon when fluorescence functional groups assembled, which is a common problem of fluorescent molecular imaging. In 2001, the Tang’s team found the aggregation-induced emission(AIE) phenomenon, refers to AIE, the fluorescence groups are practically nonemissive in solution states, while boosts their luminescence efficiency dramatically in solid or gathered condition, means that use this kind of fluorescence groups as fluorescence probe have an incomparable advantage compared with traditional fluorescent probe. But at present, about the cell dyeing sites and the influence of cell growth activity and apoptosis are still unclear. In the compounds with AIE properties, tetraphenylethene(TPE) and its derivatives can emit strong blue fluorescence in gathered state, the excellent luminous performance and convenient synthesis make TPE and its derivatives as probes to be an important component in biological detection field.Objective Research TPE’s cell dyeing sites and detect its effect on cell growth activity and cell apoptosis respectively in tumor cells and normal cells.Methods Using MEM cultivate human cervical cancer cell line HeLa and DMEM cultivate human fibroblast HMF. Adopt McMurry reaction synthesis TPE and detect its AIE characteristics. TPE diluted into different concentrations, then add into the medium for culture cells and test different fluorescence intensity, all images analysis using NIS Elements F software. Using WST?-8 method to detect cell growth activity and Annexin V-FITC/PI method to detect cell apoptosis, all measurement datas are testing by the SPSS13.0 for Windows software.Results TPE is completely dissolving in DMSO. Under the UV illumination, TPE is non-emissive in solution state, but emit strong blue light in solid state, with TPE concentrations increasing the fluorescence intensity is strengthening. The blue fluorescence mainly locate in cytoplasm, nuclear is not shading, fluorescence intensity and TPE concentration become direct ratio. Tumor cells and normal cells growth activities are all more than 90% after TPE treatment. Using flow cytometry detect apoptosis rate of HeLa cells and HMF cells, compared with controls, the differences between 10μmol/L group and 20μmol/L group are no statistically significant. (P > 0.05)Conclusions Based on this, the TPE could be modified and used as label for organelles or certain molecules in live cell with high selectivity. Therefore, TPE will have a great perspective in biomedicine.

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