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dc.contributor.advisorDholakia, Kishan
dc.contributor.authorMthunzi, Patience
dc.coverage.spatial255en_US
dc.date.accessioned2010-11-08T13:04:09Z
dc.date.available2010-11-08T13:04:09Z
dc.date.issued2010-11-30
dc.identifier.urihttps://hdl.handle.net/10023/1254
dc.description.abstractRecently, laser light sources of different regimes have emerged as an essential tool in the biophotonics research area. Classic applications include, for example: manipulating single cells and their subcellular organelles, sorting cells in microfluidic channels and the cytoplasmic delivery of both genetic and non-genetic matter of varying sizes into mammalian cells. In this thesis several new findings specifically in the optical cell sorting as well as in the photo-transfection study fields are presented. In my optical cell sorting and guiding investigations, a new technique for enhancing the dielectric contrast of mammalian cells, which is a result of cells naturally engulfing polymer microspheres from their environment, is introduced. I explore how these intracellular dielectric tags influence the scattering and gradient forces upon these cells from an externally applied optical field. I show that intracellular polymer microspheres can serve as highly directional optical scatterers and that the scattering force can enable sorting through axial guiding onto laminin coated glass coverslips upon which the selected cells adhere. Following this, I report on transient photo-transfection of mammalian cells including neuroblastomas (rat/mouse and human), embryonic kidney, Chinese hamster ovary as well as pluripotent stem cells using a tightly focused titanium sapphire femtosecond pulsed laser beam spot. These investigations permitted advanced biological studies in femtosecond laser transfection: firstly, the influence of cell passage number on the transfection efficiency; secondly, the possibility to enhance the transfection efficiency via whole culture treatments of cells thereby, synchronizing them at the mitotic (M phase) as well as the synthesis phases (S phase) of the cell cycle; thirdly, this methodology can activate the up-regulation of the protective heat shock protein 70 (hsp70). Finally, I show that this novel technology can also be used to transfect mouse embryonic stem (mES) cell colonies and the ability of differentiating these cells into the extraembryonic endoderm.en_US
dc.language.isoenen_US
dc.publisherUniversity of St Andrews
dc.relationIEEE Journal of Selected Topics in Quantum Electronics, Vol 16 (3), May/June, 2010en_US
dc.relationJournal of Biomedical Optics, Vol 15 (4), July/August, 2010en_US
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/
dc.subjectOptical cell sortingen_US
dc.subjectIntracellular dielectric taggingen_US
dc.subjectAxial and scattering forcesen_US
dc.subjectPhoto-transfectionen_US
dc.subjectFemtosecond laser pulsesen_US
dc.subjectCell arrestingen_US
dc.subjectNeuroblastomasen_US
dc.subjectPluripotent stem cellsen_US
dc.subjectEmbryonic stem cell differentiationen_US
dc.subject.lccQH585.5F56M8
dc.subject.lcshFlow cytometryen_US
dc.subject.lcshFemtosecond lasersen_US
dc.subject.lcshTransfectionen_US
dc.subject.lcshMammals--Cytologyen_US
dc.titleOptical sorting and photo-transfection of mammalian cellsen_US
dc.typeThesisen_US
dc.contributor.sponsorCouncil for Scientific and Industrial Research (CSIR), South Africaen_US
dc.contributor.sponsorUniversity of St Andrewsen_US
dc.type.qualificationlevelDoctoralen_US
dc.type.qualificationnamePhD Doctor of Philosophyen_US
dc.publisher.institutionThe University of St Andrewsen_US


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Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
Except where otherwise noted within the work, this item's licence for re-use is described as Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported