HOPE-SIM, a cryo-structured illumination fluorescence microscopy system for accurately targeted cryo-electron tomography

Li, Shuoguo; Jia, Xiaoyang; Niu, Tongxin; Zhang, Xiaoyun; Chen, Qi; Xü, Wei; Deng, Hongyu; Sun, Fei; Ji, Gang

doi:10.1038/s42003-023-04850-x

Cited by 4 publications

(2 citation statements)

References 59 publications

(81 reference statements)

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“…By juxtaposing findings from complementary techniques, researchers can validate their results, offering a holistic understanding of cellular events. Fluorescence microscopy, for example, is always used as an initial step, which is then followed by in-cell NMR or cryo-EM experiments. ,,, This sequence ensures that target proteins are accurately localized within the cell. Subsequently, when this information is combined with in-cell NMR/Cryo-EM data, it can reveal the components with which the target proteins might interact.…”

Section: Discussionmentioning

confidence: 99%

Advanced Techniques for Detecting Protein Misfolding and Aggregation in Cellular Environments

Bai,

Zhang,

Dong

et al. 2023

Chem. Rev.

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Protein misfolding and aggregation, a key contributor to the progression of numerous neurodegenerative diseases, results in functional deficiencies and the creation of harmful intermediates. Detailed visualization of this misfolding process is of paramount importance for improving our understanding of disease mechanisms and for the development of potential therapeutic strategies. While in vitro studies using purified proteins have been instrumental in delivering significant insights into protein misfolding, the behavior of these proteins in the complex milieu of living cells often diverges significantly from such simplified environments. Biomedical imaging performed in cell provides cellular-level information with high physiological and pathological relevance, often surpassing the depth of information attainable through in vitro methods. This review highlights a variety of methodologies used to scrutinize protein misfolding within biological systems. This includes optical-based methods, strategies leaning on mass spectrometry, in-cell nuclear magnetic resonance, and cryo-electron microscopy. Recent advancements in these techniques have notably deepened our understanding of protein misfolding processes and the features of the resulting misfolded species within living cells. The progression in these fields promises to catalyze further breakthroughs in our comprehension of neurodegenerative disease mechanisms and potential therapeutic interventions.

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Section: Discussionmentioning

confidence: 99%

Advanced Techniques for Detecting Protein Misfolding and Aggregation in Cellular Environments

Bai,

Zhang,

Dong

et al. 2023

Chem. Rev.

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“…For example, phase changes between growth and shrinkage of microtubules have been observed in this manner. [ 57 ] Since then, the use of cryo‐CLEM has vastly expanded (Figure 1B) and many groups started to work on enhanced localization of areas of interest (e.g., through correlative lamellae preparation) and on closing the temporal gap between light microscopic observation, cryo‐arrest and cryo‐EM using rapid transfer and in situ techniques [ 58–60 ] Without these, however, dynamic cryo‐CLEM is limited to relatively slow processes with typical timescales in the range of minutes with well‐characterized morphologies. b) Rapid sample transfer : The first approaches to not only correlate, but to capture specific events occurring live in the light microscope focused on rapid sample transfer by moving the sample from the light microscope quickly to a cryofixation device. The rapid transfer system developed by Verkade et.…”

Section: Dynamic Clem Techniquesmentioning

confidence: 99%

Time Resolved Cryo‐Correlative Light and Electron Microscopy

Szabo,

Burg

2024

Adv Funct Materials

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Complex materials exhibit fascinating features especially in situations far from equilibrium. Thus, methods for investigating structural dynamics with sub‐second time resolution are becoming a question of interest at varying spatial scales. With novel microscopy techniques steadily improving, the temporal and spatial limits of multiple imaging methods are investigated with an emphasis on the important role of correlative imaging and cryo‐fixation. A deep‐dive is taken into cryo‐correlative light and electron microscopy (CLEM) as a starting point for multimodal investigations of ultrastructural dynamics at high spatiotemporal resolution. The focus is on highlighting the different microscopy methods that capture the following key aspects: 1) samples are as close to native state as possible 2) dynamic process information is captured, 3) high structural resolution is enabled. Additionally, the size of samples that can be imaged under these conditions is looked at and approaches not only focusing on single molecules, but larger structures are highlighted.

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