Limitations to light microscope resolution in biological preparations and methods for enhanced resolution using fabricated microstructures

McIntyre, Stuart (2013) Limitations to light microscope resolution in biological preparations and methods for enhanced resolution using fabricated microstructures. PhD thesis, University of Glasgow.

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Abstract

Fluorescence imaging has become a vital tool for understanding cardiac cell function. With the advent of calcium-sensitive and fast response voltage-sensitive dyes, it is now possible to gain complex physiological recordings from various cardiac muscle preparations, while simultaneously optically sectioning the samples. However, due to the complexity of these experimental set-ups, it is difficult to set up control samples that allow the user to predict how their sample is affecting the quality of the data being captured. The aim of this thesis is to assess the strengths and weaknesses of the different types of optical sectioning microscope methods that are currently available in many biological labs. These include confocal, two-photon and selective illumination microscopes . The resolution capabilities of these systems and the associated objective lenses were investigated to determine the optimum conditions for their use. This work includes investigations of the contribution of signal noise and performance of motorised z-stage systems used in these microscope systems to the ability of assess optical resolution.. The optical components of cardiac muscle are investigated individually. This includes the scattering properties of the myocardium and the effects of the refractive index mismatch, as well as the intrinsic fluorescent substances found in cardiac muscle. Microstructures are used to generate novel ways of overcoming some of the problems faced when imaging cardiac muscle in order to improve the sensitivity and resolution of these systems. Finally, a method is investigated to enhance the the relative concentration of live cardiac:dead cells in after dissociation from intact hearts. This approach would aid the study of isolated cardiac cells be reducing the interference from damaged cells.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Subjects: R Medicine > RC Internal medicine
T Technology > T Technology (General)
Colleges/Schools: College of Science and Engineering > School of Engineering > Biomedical Engineering
Funder's Name: UNSPECIFIED
Supervisor's Name: Smith, Prof. Godfery and Cooper, Prof. Jon
Date of Award: 2013
Depositing User: Mr Stuart Mcintyre
Unique ID: glathesis:2013-4245
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 14 May 2013 11:41
Last Modified: 14 May 2013 11:43
URI: http://theses.gla.ac.uk/id/eprint/4245

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