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|Title: ||Advanced photonic methodologies for the 'in vitro' manipulation of cellular systems|
|Authors: ||McDougall, Craig|
|Supervisors: ||Brown, Tom|
|Issue Date: ||2011|
|Abstract: ||This thesis investigates the application of a variety of optical techniques for the
manipulation of single cells and their local micro-environment. The methodologies
developed provide enhanced control over a single cell under study affording exquisite
spatial and temporal control over biological processes of interest. The work presented
within the thesis can be split into three distinct categories.
The first of these provides an investigation in light activated “caged” molecular
probes. This work generated several new compounds which were then applied to
providing control over processes involved in pain, mitochondrial intracellular signalling and memory processes in the central nervous system. Application of caged
neurotransmitters then demonstrates the first in vitro wavelength orthogonal photolysis of biologically relevant substances. Such a technique has great potential in
the study of fundamental interactions within the processes underpinning memory
and cognitive function.
Secondly the application of optical injection techniques for the introduction of
membrane impermeable species of interest is presented. An exploration of laser
sources and optical systems has yielded two new strategies for optical injection.
The targeted introduction of fluorescent stains, nucleic acids and gold nanoparticles
to the interior of live mammalian cells demonstrates the power of these techniques.
Thirdly, an investigation in optical trapping and optical injection provides simplified micromanipulation techniqes for application to biological studies. The use
of capillaries as reservoirs for reagents of interest has realised a procedure for the
reduction of large-scale chemical assays to a single cell level in static flow. When
this technique is combined with intelligent control over the trapping laser source’s
temporal behaviour, the interaction with the sample under study can be tailored for biological amiability or sample ablation. In this way a single laser source can be employed for the optical trapping and nanosurgery of a biological sample. A final study
is presented demonstrating initial results for the targeted optical injection of caged
compounds into mammalian cells. This methodology draws on the strengths of optical injection and caging technologies and presents a significant step forward in the
level of control afforded over a biological system under study by optical techniques.
The studies presented highlight the level of control and flexibility afforded by the
application of optical manipulation and excitation strategies. Such optical methodologies extend the photonic tools available for enhanced studies in the life sciences.|
|Publisher: ||University of St Andrews|
|Appears in Collections:||Physics & Astronomy Theses|
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