eprintid: 10189770 rev_number: 12 eprint_status: archive userid: 699 dir: disk0/10/18/97/70 datestamp: 2024-04-25 11:10:13 lastmod: 2024-04-25 11:10:13 status_changed: 2024-04-25 11:10:13 type: thesis metadata_visibility: show sword_depositor: 699 creators_name: Ing, Gabriel title: Development of Liquid-phase Transmission Electron Microscopy for the study of Proteins and Peptides ispublished: unpub divisions: UCL divisions: B04 divisions: C06 divisions: F56 keywords: Electron Microscopy, Liquid-phase Electron Microscopy, Transmission Electron Microscopy note: 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. 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. date: 2024-03-28 date_type: published oa_status: green full_text_type: other thesis_class: doctoral_open thesis_award: Ph.D language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2262556 lyricists_name: Ing, Gabriel lyricists_id: GRBIN74 actors_name: Ing, Gabriel actors_id: GRBIN74 actors_role: owner full_text_status: public pagerange: 1-239 pages: 239 institution: UCL (University College London) department: Chemistry thesis_type: Doctoral citation: 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 document_url: https://discovery.ucl.ac.uk/id/eprint/10189770/2/Thesis_with_corrections_GI_3.pdf