Application of optical coherence tomography in investigating cell migration

Rey, Sara 2012. Application of optical coherence tomography in investigating cell migration. PhD Thesis, Cardiff University. Item availability restricted.

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Abstract

Chemotaxis and cell migration are important processes for life, involved in organism development and homeostasis and implicated in a number of disease states. Dictyostelium discoideum, an amoeba, is a useful model for investigation of chemotaxis and development, due to its ability to undergo chemotactic aggregation and development upon starvation. Although cell migration has been well described on planar transparent surfaces, it is uncertain how well these conditions replicate the natural environment of a cell. However, attempts to better replicate these environments generally make use of opaque substrates and 3D matrices, in which it is more challenging to image cell migration. Protocols were developed to enable optical coherence tomography, a 3D structural imaging technique which requires no sample processing or staining, to be successfully employed in imaging Dictyostelium cell migration in time-lapse on non-transparent substrata and within an agarose gel. I compared the effects of two substrates, a nitrocellulose filter and a polystyrene Petri dish on aggregating cells and found differences in speed but not persistence. Extension of this to include cells within agarose revealed that these cells exhibited less directed migration, but their velocity was unaffected. I showed that cells lacking myosin II failed to complete development within an agarose gel and had significantly reduced velocity and directional migration when compared to their parent strain. Furthermore, the velocities of cells migrating within agarose gel were bimodally distributed, potentially indicating two distinct cell populations, fast and slow, and fast movement was shown to be largely myosin II dependent. Great potential therefore exists for cell-substrate and cell-matrix interactions to affect the migration character of cells, even those, such as Dictyostelium, which do not form strong focal adhesions. Moreover a properly ordered cytoskeleton is implicated in enabling cells to effectively utilise different modes of cell motility.

Item Type:	Thesis (PhD)
Status:	Unpublished
Schools:	Schools > Biosciences
Subjects:	Q Science > QH Natural history > QH301 Biology R Medicine > RE Ophthalmology
Uncontrolled Keywords:	Optical Coherence Tomography ; noninvasive imaging ; cell migration ; chemotaxis ;, 4D imaging ; 3D imaging ; myosin II ; Dictyostelium discoideum
Funders:	Biotechnology and Biological Sciences Research Council
Date of First Compliant Deposit:	30 March 2016
Last Modified:	10 Oct 2017 14:57
URI:	https://orca.cardiff.ac.uk/id/eprint/44209

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