Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy
<p>The advent of single-particle tracking and super-resolution imaging techniques have brought forth a revolution in the field of single-molecule optical microscopy. This thesis details the development and subsequent implementation of the technique known as interferometric scattering microscop...
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2015
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| _version_ | 1826286337626472448 |
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| author | Arroyo, J |
| author2 | Kukura, P |
| author_facet | Kukura, P Arroyo, J |
| author_sort | Arroyo, J |
| collection | OXFORD |
| description | <p>The advent of single-particle tracking and super-resolution imaging techniques have brought forth a revolution in the field of single-molecule optical microscopy. This thesis details the development and subsequent implementation of the technique known as interferometric scattering microscopy as a novel single-molecule tool to study nanoscopic dynamics and their underlying potentials. Specifically, Chapter 2 lays out the theoretical framework and draws comparisons between this technique and other state-of-the-art single-molecule optical approaches. Chapter 3 provides a detailed description for the design and implementation of an interferometric scattering microscope including alignment, instrumentation, hardware interfacing, image processing and respective characterisation to achieve the highest levels of performance. The following two chapters use model systems, namely the diffusion of receptor GM1 in a supported lipid bilayer and the movement of molecular motor myosin 5a, to demonstrate the intrinsic shot-noise limited nature of the technique, its ability to decouple the temporal resolution from localisation precision, and highlight the importance of taking both parameters into consideration when drawing conclusions about the dynamics of each model system. Chapter 6 provides a proof-of-concept study on the limits of sensitivity and demonstrates for the first time the all-optical label-free imaging, detection and tracking of a single protein. In the last chapter, interferometric scattering microscopy is used to quantitatively study dynamic heterogeneous systems in situ at the single particle level and thus serves as a proof of principle for future label-free studies beyond the realms of biophysics.</p> |
| first_indexed | 2024-03-07T01:42:16Z |
| format | Thesis |
| id | oxford-uuid:97464d98-5b5a-42a9-9d02-19b15b7b9b57 |
| institution | University of Oxford |
| last_indexed | 2024-03-07T01:42:16Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | oxford-uuid:97464d98-5b5a-42a9-9d02-19b15b7b9b572022-03-26T23:58:24ZInvestigation of nanoscopic dynamics and potentials by interferometric scattering microscopyThesishttp://purl.org/coar/resource_type/c_db06uuid:97464d98-5b5a-42a9-9d02-19b15b7b9b57Biophysics--TechniqueORA Deposit2015Arroyo, JKukura, P<p>The advent of single-particle tracking and super-resolution imaging techniques have brought forth a revolution in the field of single-molecule optical microscopy. This thesis details the development and subsequent implementation of the technique known as interferometric scattering microscopy as a novel single-molecule tool to study nanoscopic dynamics and their underlying potentials. Specifically, Chapter 2 lays out the theoretical framework and draws comparisons between this technique and other state-of-the-art single-molecule optical approaches. Chapter 3 provides a detailed description for the design and implementation of an interferometric scattering microscope including alignment, instrumentation, hardware interfacing, image processing and respective characterisation to achieve the highest levels of performance. The following two chapters use model systems, namely the diffusion of receptor GM1 in a supported lipid bilayer and the movement of molecular motor myosin 5a, to demonstrate the intrinsic shot-noise limited nature of the technique, its ability to decouple the temporal resolution from localisation precision, and highlight the importance of taking both parameters into consideration when drawing conclusions about the dynamics of each model system. Chapter 6 provides a proof-of-concept study on the limits of sensitivity and demonstrates for the first time the all-optical label-free imaging, detection and tracking of a single protein. In the last chapter, interferometric scattering microscopy is used to quantitatively study dynamic heterogeneous systems in situ at the single particle level and thus serves as a proof of principle for future label-free studies beyond the realms of biophysics.</p> |
| spellingShingle | Biophysics--Technique Arroyo, J Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title | Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title_full | Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title_fullStr | Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title_full_unstemmed | Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title_short | Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| title_sort | investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy |
| topic | Biophysics--Technique |
| work_keys_str_mv | AT arroyoj investigationofnanoscopicdynamicsandpotentialsbyinterferometricscatteringmicroscopy |