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New approaches to the study of biophysicochemical processes

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Meadows, Katherine E. (2013) New approaches to the study of biophysicochemical processes. PhD thesis, University of Warwick.

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Abstract

This thesis is concerned with the study of biophysicochemical processes using electrochemistry and related techniques. The first part of the thesis discusses the electrochemical detection of biological species, and characterisation of the electrode materials employed. A comparison of two novel forms of carbon electrode, namely carbon nanotubes and polycrystalline boron doped diamond (pBDD), with more conventional carbon electrode materials reveals their enhanced characteristics for bioelectrochemistry, with improved sensitivity and resistance to fouling. These materials are further characterised using novel high-resolution electrochemical imaging methods, to determine heterogeneous electron transfer rates for a number of different redox species. The kinetic rate constants are determined from measured electrochemical currents using finite element method (FEM) modelling, which proves to be a powerful technique for the quantitative analysis of intrinsic system parameters that cannot be studied directly. The electrochemical response of isolated regions of pristine SWNTs is investigated using scanning electrochemical cell microscopy, demonstrating high electrochemical activity at the nanotube sidewalls. A similar analysis of the different facets of pBDD is performed using intermittent contact scanning electrochemical microscopy coupled with FEM simulations, revealing that the electroactivity is strongly influenced by the local density of states of the material.

New techniques are also presented for the investigation of transport processes at membrane interfaces. A new method of bilayer formation is developed, which overcomes many of the limitations of current techniques, and is used to investigate the permeation rates of a series of aliphatic carboxylic acids. Using confocal laser scanning microscopy (CLSM) with a pH-sensitive uorophore, the pH change as a weak acid permeates across the bilayer can be visualised, and the permeation coefficient determined by comparison with FEM simulations. This reveals a trend of increasing permeability with lipophilicity. Finally, CLSM is used to study the lateral diffusion of protons at lipid bilayers and other surfaces.

Protons are generated galvanostatically by a UME positioned close to the substrate, altering the local pH which can be visualised by means of a pH-sensitive uorophore. The uorescence profile is again compared to FEM simulations, allowing the lateral diffusion coefficient to be determined.

Item Type: Thesis (PhD)
Subjects: Q Science > QC Physics
Q Science > QD Chemistry
Q Science > QH Natural history > QH301 Biology
Q Science > QP Physiology
Library of Congress Subject Headings (LCSH): Biophysics, Electrochemical analysis, Bioelectrochemistry, Finite element method
Official Date: May 2013
Dates:
DateEvent
May 2013Submitted
Institution: University of Warwick
Theses Department: Molecular Organisation and Assembly in Cells
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Unwin, Patrick R.; Macpherson, Julie V.
Extent: xvii, 201 leaves : illustrations.
Language: eng

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