@phdthesis{9673dae35a3d4544a48edcd088c72625,
title = "Imaging Life at the Molecular Scale using Electrons",
abstract = "Cryogenic electron microscopy (cryo-EM) has become a powerful technique to understand the structure and function of proteins. Thanks to substantial technical advancements in the 2010s, the frequent determination of atomic structures is now possible. This thesis encompasses chapters that contribute to both the remaining technical challenges of cryo-EM, and to the advancement of biological insight through the application of cryo-EM to a motility machinery called a gas vesicle. As an overarching theme, the thesis is titled {\textquoteright}Imaging Life at the Molecular Scale using Electrons{\textquoteright}. Chapter 1 provides an introduction into methods to study structure and function in biology at the molecular scale, and into the development of cryo-EM since its inception. A comprehensive review of previous structural work on gas vesicles follows.Technological improvements in electron detectors have significantly enhanced the quality of protein images obtainable. However, predicting the success of cryo-EM structure determination still poses a challenge, as it is heavily dependent on the unique properties of the sample and its behavior during the sample preparation process. Proteins are typically forced into an thin liquid layer, which is frozen by quick immersion into a cryogen. The large air-water interface can cause proteins to aggregate, denature or adopt preferred orientation, hindering structure determination. The thickness of the layer is also hard to control - too thick layers lead to poor image contrast, too thin layers damage the protein.....",
keywords = "cryo-EM, sample preparation, gas vesicle, time resolution",
author = "S. Huber",
year = "2024",
doi = "10.4233/uuid:9673dae3-5a3d-4544-a48e-dcd088c72625",
language = "English",
type = "Dissertation (TU Delft)",
school = "Delft University of Technology",
}