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The Detection of Active Small Molecules Substances in the Cell Baseg on Circle Amplification

Author: FuYouJian
Tutor: ZuoBin
School: Qingdao University of Science and Technology
Course: Organic Chemistry
Keywords: nano particles fluorescence electrochemiluminesce(ECL) APS GSH tumour cell
CLC: R73-3
Type: Master's thesis
Year: 2013
Downloads: 4
Quote: 0
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Abstract


In the chapter one, first of all, the formation of tumor cells and detection was introduced briefly, then introduced the fluorescence, chemiluminescence and electrochemiluminescence. The analyzing of previous electrochemiluminescence and fluorescence, we know its basic principle, tendency of the current detection and the detection level of the current.In the chapter two, we prepared the luminol gold nanoparticle (luAu NPs), due to its particularity properties, this kind of gold nanoparticles have electrogenic chemiluminescence and fluorescence properties. Due to the breaking in the disulfide bond of GSH, we established the luAu NPs fluorescence detection system to detect the content of GSH and the GSH content in the cell. The detection of this system was as low as2.0×10-7M, and can detect100-1000K562cells.In the chapter three, based on the synthesis of luAu NPs, we designed a cycle amplification and APS enhanced electrochemiluminescence system to detect the tumor cells. This system not only can sensitive, specific detect the target cells, but also can be used in sensitive detection of target DNA. The method of "one-pot" in the electrode surface can gain signals quickly and easily. The APS can enhance luminol ECL signal. Optimization the implementation condition, the DNA linear range was2.0×10-16-1.0×10-13M and the detection limit was2.0×10-16M. The method is designed for analyzing cancer cell (about10-1280cells) and the detection limit is as low as10cells. This system has a good application prospect in the field of biology analysis.In the chapter four, we prepared the graphene oxide (GO), the phase and morphology of products were characterized by High resolution transmission electron microscopy. Through isothermal amplification technique and GO as the nano container to load the ssDNA and disulfide bond issue of DNA, we can detect the GSH. Optimization the implementation condition, the GSH linear range was1.0×10-10-1.0×10-7M and the detection limit was1.0×10-10M. The method is designed for analyzing cancer cell (about10-1000cells) and the detection limit is as low as10cells. This scheme has high sensitivity and high specificity, and has very important significance in the analysis of the reduced biological thiol compounds in the cancer cells.

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