| Labeled immunoassay are the total conception of a series of highly sensitive and specific examine techniques, and have been widely applied in the basic medicine and clinic field for their special advantages. With the exertion of new theories and new techniques, the labeled immunoassay have got great development Having the advantages of RIA but without its disadvantage of radioactivity, Time-resolved Fluorescence Immunoassay (TRFIA) has become one of the hotspots in the field of labeled immunoassay applicatioa So, it's very important to develop sensitive, operating easily and cheap time-resolved fluorometer for basic medicine research and clinic application.In this thesis, the several methods used widely in clinical detection were introduced firstly, and the TRFIA was introduced in detail. Then, through the theory analysis and practical experiments, the characteristics of excitation spectrum and emission spectrum of lanthanide Eu and its Eu3+ chelaes were studied, some valuable results were obtained. Based on the research of spectrum, a widely applicable semiautomatic time-resolved fluorometer was produced, which was suitable for normal time-resolved fluorescence immunoassay reagentIn the study of excitation and emission spectra of lanthanide Eu and its Eu3+ chelaes, the absorption and emission spectra was firstly analyzed from the structure of Eu3+ and the energy transition, then through experiments of different light wave, the optimized excitation and emission spectra were obtained The results of the experiments showed that the best wavelength to excite Eu3+ chelates is 300-350nm, and its fluorescence spectrum is in the range of 610nm-620nm. Based on the results from theory and experiment, the excitation mechanism was obtained. At the same time, the fluorescence lifetime of Eu3+ chelates was studied by using PMT and oscillograph. The above achievements provide valuable information for the design of time-resolved fluorometer.Based on the study of spectrum, the components and technology requirement of time-resolved fluorometer were presented And a concrete method to actualize the system of pulsed Xenon lamp and the assemblage of filter setting was developed Sound clinical evaluation project was designed At last, clinical evaluation of the time-resolved fluorometer was conducted.Results showed that the time-resolved fluorometer has many advantages such as sensitive, stable, easy to operate and low cost In conclusion, the time-resolved fluorometer we designed is an advanced and suitable clinical immunoassay set. |
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