Research Of Monitoring SH-SY5Y Cells Electrochemical Behavior Based On Film ITO Microelectrodes

As we all know, cell is the basic unit of organisms and life activities, and cell research is the premise of understanding life science.Cell metabolism, photosynthesis, the transmission of information and transmembrane transport of substances are closely related with the the transmission of cell charged particles or the electrically active particles, therefore, electrochemical reaction is the foundation of vital activities of cells, and electrochemical method is one of the most effective research methods of life activities. Due to the heterogeneity of biological tissue, there are some differences between cells, so the results of cell population analysis are often inaccurate. Single cell analysis plays an important role in the early diagnosis and treatment of disease, drug design and the control of drug side effects.ITO microelectrodes which prepared by micro-processing technology have excellent transparency, high conductivity,wide electrochemical window and low cost. Based on these advantages, the film ITO microelectrodes can be combined with fluorescence and chemiluminescence, so it has been widely used as a platform for cell research in recent years.In this paper, SH-SY5Y cells were chosen as the object of study, and homemade film ITO microelectrodes were chosen as a platform. Amperometry was used to monitor SH-SY5Y single cell exocytosis, and impedance method was used to real time monitor growth status of SH-SY5Y cells on the microelectrodes.This paper consists of the following three chapters. Chapter One:PrefaceThe cell which is the basic unit of life activities was first introduced in this chapter, then we introduced electrochemical analysis of single cell exocytosis and electrical impedance analysis of cells. We focused on the principles of exocytosis, the electrochemical technique, types of microelectrodes, the significance of single cell analysis and the purpose and significance of this paper.Chapter Two:Monitoring SH-SY5Y single cell exocytosis with microelectrodesExocytosis release specific biochemical or chemical messengers, which plays crucial role in biological process. Norepinephrine, dopamine and epinephrine are catecholamines, which have electrochemical activity, and which can be monitored by amperometric. ITO film microelectrodes which were prepared by UV lithography method owes high sensitivity, and they can meet the requirements of the single-cell exocytosis study. Compared with traditional carbon fiber microelectrodes, if we used film ITO microelectrodes to monitor exocytosis, the experimental operation was very simple. This study investigated exocytosis of different forms of SH-SY5Y cells, the spindle cells had better exocytosis activity compared with the spherical cells. Both carbon fiber microelectrodes and film ITO microelectrodes were used to monitor exocytosis, experimental results showed that the film ITO microelectrodes were able to monitor more exocytosis signals, In this experiment we also used different stimulants, and we found that there were some differences in exocytosis response time. Chapter Three: Based on film ITO micro-electrode electrochemical impedance spectroscopy monitoring the growth status of SH-SY5Y cellsFor most cells, cell adhesion is the most important initial step in cell growth, ITO conductive film glass has the advantages of low cost, transparent and stable performance.We used ultraviolet lithography method to build film ITO microelectrode array, and we real-time monitored growth status of SH-SY5Y cells with electrochemical impedance method. From the cell index curve (CI), the CI values had different degree of rise after adding cell suspension. When the concentration of anticancer drug arsenic trioxide (AS2O3) increased to40μmol/L, we could see that CI value decreased significantly in1hour, from inverted microscope cell morphology also underwent a significant change at the same time, the phenomenon indicated that changes of CI value had correlation with the changes of cell morphology, but the CI value could be able to monitor changes in cell status in the early stages of drug effects. The experimental results showed that our homemade film ITO microelectrode array can be used to monitor status of SH-SY5Y cell growth with real-time and label-free method, we also provided a good platform for future drug screening and cellular dynamics research.