Investigation of a model FRET system with multiple donors and acceptors. Allowing validation of data interpretation for studies of cells

In this study the foundations are laid for using a fluorescence microscopy system to view multiple fluorescent species in mammalian cells as a possible technique in investigating the oligomers status of GPCRs. The system utilises spectrally resolved intensity, anisotropy and lifetime measurements. All these techniques are proven to be useable and comparable to the literature. Its weaknesses such as illumination intensity are recognised and its more novel capabilities are utilised. Cho cells are imaged using a confocal microscope and the plasmid sequences providing them with fluorescence through YFP and CFP are established to be correct. In the pursuit of fine tuning the microscope system to allow the live cells images to be taken and any FRET recognised, a model system from the literature is examined. However this turns out to be a fascinating system all of its own; the calculations lead to conclusions that the FRET occurring are in fact predicting inner filter effects along with other unidentified processes. This study looks at why this is the case.

Name dc:type.qualificationname

mphil

Level dc:type.qualificationlevel

masters

Grantor dc:publisher.institution

Nottingham Trent University

Year dc:date.issued

2011

Author dc:creator

• Gumbs, WGA

Language dc:language

en