Determination Of Glutathione In Single Cell Based On Capillary Electrophoresis With Electrocehmiluminescence Detection

In the chapter I, at first the basic principles of capillary electrophoresis and its development were introduced briefly. Then the appliance of capillary electrophoresis in single-cell analysis (including single-cell sampling, analysis detection methods of single cell, and the techniques of lysing single cell respectively), the work for capillary electrophoresis with electrochemiluminescence detection was summarized , in the end immobilization and application of electrochemiluminescence sensor were introduced.In the chapter II, we used capillary electrophoresis-electrochemiluminescence (ECL) to detect the content of glutathione in rat peritoneal mast cells based on tris(2,2'-bipyridine) ruthenium(II) (Ru(bpy)₃²⁺) which is the concentration of 4.00×10^-3 mol-L^-1, The optimum conditions of separation and detection are 5.00×10^-2 mol-L^-1 NaH2PO4- Na2HPO4 (pH 7.5) for the buffer solution, 12 kV for the separation voltage, 10 kV and 10 s for the injection voltage and the injection time, and 1.2 V (vs. Ag/AgCl) for the detection potential. The amount of glutathione (GSH) in individual rat peritoneal mast cell on average is 18.7 fmol.In the chapter III, the method of capillary electrophoresis with electrochemiluminescence deteaion was used to detect the content of glutathione in single cells. The detected content of glutathione in individual rat peritoneal mast cells was from 9.30 to 25.9 fmol. The content of single B lymphoma cells (Ramos cell) is from 2.6 to 6.5 fmol and the amount of single human cervical cancer cells (Hela cell) is from 24.3 to 57.2 fmol.In the chapter IV, several kinds of methods to fix Ru(bpy) 32 + on the carbon fiber micro-disk bundle electrode surface was attempted, in the end we made the micro-electrode which was immobilized Ru(bpy)₃²⁺ in carbon nanotubes (CNT)/Nafion composite film onto the carbon fiber micro-disk bundle electrode. Its electrochemiluminescence was strong and stability. We studied its application in capillary capillary electrophoresis preliminary.In the chapter V, the device of single-cell electroporation apparatus with the single-cell electroporation injector was made. The effect of electric pulse for the cell was studied initially during the progress of electroporation.