Thesis
Development of computational methods for standing wave microscopy
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2019
- Thesis identifier
- T15380
- Person Identifier (Local)
- 201572756
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Optical microscopy is an important tool in biomedical sciences and allows for the imaging of single cells, and has become vital and versatile tool for understanding cellular and subcellular processes. However, due to the wave nature of light, the spatial resolution is limited. As a result, most of the important biological process of a cell take place below 200 nm, which is beyond the resolution capabilities of conventional optical microscopy techniques. In this work, I used multi-planar standing wave microscopy, an axial super resolution technique, where incident and reflected light from a mirror surface, create a standing wave. Multiple planes of lights are generated with a spacing of λ/2n, with an axial resolution of λ/4n, where λ is the excitation wavelength of light and n is the refractive index. When the multiple planes of light intersect a fluorescently labelled specimen, the result is an image with 3D information encoded in a 2D image. I showed that multi-planar standing wave images of healthy and Plasmodium berghei infected red blood cells had clear morphological deformations in the cell membrane. Furthermore, by applying a combination of noise filtering and segmentation techniques, 2D anti-nodal plane information was extracted. Next, I applied a priori knowledge to the standing wave red blood cells images to create a 3D reconstruction of the bottom concave surface. To overcome, the limitations of single-wavelength standing wave (SW) microscopy a new method, which we call TartanSW multi-excitation, was applied to biological specimens. This reduced the modulation gap when compared with the use of a single-excitation wavelength. Furthermore, TartanSW multi-excitation and multi-emission both showed a unique spectral signature that reduced the ambiguity in the geometry of the specimen. Lastly, by utilising the phase difference between the different TartanSW multi-excitation wavelengths, I showed that in principle, the phase difference could be used to determine the axial height information from a 2D encoded standing wave pattern.
- Advisor / supervisor
- McConnell, Gail
- Resource Type
- Note
- This thesis was previously held under moratorium from 27th November 2019 until 27th November 2024.
- DOI
- Date Created
- 2019
- Former identifier
- 9912772693402996
Relations
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PDF of thesis T15380 | 2021-07-02 | Public | Download |