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Development of Liquid-phase Transmission Electron Microscopy for the study of Proteins and Peptides

Ing, Gabriel; (2024) Development of Liquid-phase Transmission Electron Microscopy for the study of Proteins and Peptides. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

Liquid-phase transmission electron microscopy (LTEM) is a powerful technique to study the dynamics of molecular processes occurring in real-time. While the technique is becoming increasingly established in inorganic and metal-containing systems, the next frontier of LTEM is the study of organic and biological material at the molecular level. In this project, I have worked on establishing LTEM as an effective imaging method to image biomolecules. To this end, I have imaged several control proteins including ferritin, β-galactosidase and apoferritin, with a variety of state-of-the-art LTEM systems. This project has proved highly challenging due to a combination of low inherent contrast, radiation sensitivity of the proteins and fast motion of the particles. Alongside the imaging of single proteins, an applied study of the aggregation of amyloid-β peptide was also performed, yielding interesting data on the nucleation of amyloid-β oligomers and fibrils, as well as demonstrations of phase separation. The work on amyloid-β demonstrates the advantages of imaging large-scale systems with LTEM, despite the challenges associated with imaging individual proteins. Finally, this thesis also details the development of the Python library SimpliPyTEM to aid with image processing and analysis of electron microscopy images and videos. Overall, the work herein details the development of LTEM as a method for imaging biological molecules, alongside a discussion of the limitations and future of the technique.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Development of Liquid-phase Transmission Electron Microscopy for the study of Proteins and Peptides
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Copyright © The Author 2023. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request.
Keywords: Electron Microscopy, Liquid-phase Electron Microscopy, Transmission Electron Microscopy
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry
URI: https://discovery.ucl.ac.uk/id/eprint/10189770
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