Bacciu, Antonella (2013) Sviluppo di un nuovo sistema microfluidico-amperometrico per lo studio della secrezione di dopamina in cellule PC12 utilizzando un microsensore nanostrutturato. Doctoral Thesis.
In this study we developed a microfluidic device for the detection of dopamine (DA) secreted by PC12 cells using microdialysis and constant potential amperometry (CPA). The PC12 cells are used as an in vitro model for the study of age-related diseases associated to an impairment of the movement consequent to a deficit of DA (i.e. Parkinson's disease). The proposed system has been optimized through the use of nanostructured microsensors based on multiwall carbon nanotubes (MWCNT) with a diameter of 15 nm and a length of about 10 µm. The nanotubes are allotropes of carbon with a cylindrical structure, are efficient thermal and electrical conductors and have huge reactive surfaces; for these characteristics they are used as transducers of electrochemical signals. The MWCNT were solubilized and deposited on top of epoxy-based graphite sensors or directly incorporated in the resin itself. The nanostructured microsensors have been calibrated with different concentrations of DA. We conducted several experiments for detecting the secretion of DA induced by drugs such as nicotine and KCl. This system is a rapid and reliable model for the study of the effects of certain drugs on the secretion of DA in vitro. The proposed model may be used for studying different cell types and the DA sensor may be easily replaced with different microsensors or biosensors that allow the in vitro detection of other important molecules, such as nitric oxide, glutamate, glucose and lactate.
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