Optical microscopy has long been an established tool in the biomedical sciences, being the preferred choice in the study of single cells and tissue sections. The realisation of the confocal laser scanning microscope in the 1980s led to major advances in the way optical microscopy is implemented, paving the way for the use of interference techniques such as 4Pi microscopy to increase the optical resolution, and for nonlinear microscopy techniques such as two-photon microscopy, which allows deeper penetration and the imaging of live specimens as a consequence of reduced photo-bleaching, and coherent anti-Stokes Raman scattering (CARS) microscopy, which produces high-contrast images without the need for fluorescent staining. In this work, I discuss advances in nonlinear and interference techniques available for biomedical imaging. I present a simultaneous near-field and far-field viewer for use in aligning the input beams in a CARS microscope and in a sum-frequency-generation- based two-photon microscope. I show 3D optical sectioning of whole mouse embryos using the Mesolens, a giant microscope objective capable of subcellular resolution in a 5 mm field of view, and present theoretical calculations on its use for two-photon microscopy. I present fast recording of synaptic events in neurones, with reduced photo-bleaching, using widefield two-photon microscopy. Finally, I show multiple super-resolved sections are obtained using a laser scanning standing wave microscope, generating precise contour maps of the surface membrane of red blood cells and revealing 3D information from a single image.
|Date of Award||29 May 2015|
- University Of Strathclyde
|Sponsors||University of Dundee & University of Strathclyde|
|Supervisor||Gail McConnell (Supervisor) & Erling Riis (Supervisor)|