.
Centre for Cell Engineering, University of Glasgow
A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the University of Glasgow.
October 2000.
Copyright (c)2000 Carolyn McEwan. All rights reserved.
Email: carolyn@lochan.org.
Since the end of the last century, microelectrodes have been used as a powerful technique for investigating the nervous system and its properties. However many of the techniques employed disrupt the cell membrane, and ultimately cause cell death. Planar microelectrode arrays have the potential to provide a relatively physiological method of stimulating neurons.
A procedure is described whereby a human neuroblastoma cell line, SH-SY5Y, may be stimulated by planar microelectrode arrays, and any response generated viewed optically. The project focussed initially on optimising cell culture conditions on the arrays in order to provide a healthy environment, which would in turn promote electrical activity of the neurons. The design of the planar microelectrode arrays, and the fabrication procedures used, were also modified to this end.
The cellular response to stimulation was measured using the voltage-sensitive fluorescent dye, fluo-3 AM. Fluorescent activity was recorded in one of two ways, either visually using video-imaging equipment, or as the change in voltage across a resistor in series with a photodiode. Calcium activity was generated in the cells upon application of a series of electrical pulses from the planar microelectrode arrays. This response could be increased by the application of a calcium agonist to the cells prior to stimulation.
To download the thesis:
A single 2196K zip file containing all the chapters, or
If you don't have it already, download Acrobat
Reader (for viewing PDF files) for free here:.
(this is the final version as printed on 19 June, 2001)