Thesis Abstract and Dissertation Abstracts
Microsamplinh and On-Line Separation Techniques for Mass Spectrometry: Analysis of Single Fungal Cells
Abstract Category: Science
Course / Degree: PhD
Institution / University: University of Edinburgh, United Kingdom
Published in: 2006
A macroscopic view of an organism does not reflect the extraordinary diversity of chemical and biochemical reactions occurring at the microscopic level. The biochemical analysis of a compound in a biological sample commonly involves extracting and purifying the compound from a heterogeneous mixture of cells, and provides only integrated measurements of concentrations or activities without any information at the single cell level. My research during this interdisciplinary investigation primarily focused on the development of novel single cell sampling and analysis methods applied to fungal hyphae. Micromanipulation, microsampling and mass spectroscopic analysis techniques were used to determine the concentration of biologically relevant molecules (e.g. trehalose) from living fungal hyphae, and to study the uptake of externally applied compounds (e.g. the fungicide azoxystrobin). Techniques for microsampling from single hyphae of the model organism, Neurospora crassa, and the transfer of the samples from the microscope to the mass spectrometer, following on-line separation, were developed. Capillary electrophoresis electrospray mass spectrometry (CE-ESI-MS) was used to measure intracellular concentrations of trehalose quantitatively. Methods were developed to detect external contamination of the samples taken from hyphae of Neurospora crassa, by using laser induced fluorescence (LIF). The concentration of intracellular disaccharide was measured as 1.26.+/- 0.16 mM. Using similar techniques, the intracellular concentration of the fungicide azoxystrobin was found to be 11.2 +/- 2.0 uM 5 min after being applied externally to the hyphae at a concentration of 24.8 uM.
Thesis Keywords/Search Tags:
CE, Fungal, single cell analysis, pL sample volumes, Mass spectrometry, Fungicide
This Thesis Abstract may be cited as follows:
Goodwin, R. J. A. Goodwin (2006) PhD Thesis, University of Edinburgh
Submission Details: Thesis Abstract submitted by Richard J. A. Goodwin from United Kingdom on 07-May-2007 13:00.
Abstract has been viewed 2476 times (since 7 Mar 2010).
Richard J. A. Goodwin Contact Details: Email: rjagoodwin@gmail.com
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