دورية أكاديمية
Imaging intracellular components in situ using super-resolution cryo-correlative light and electron microscopy.
| العنوان: | Imaging intracellular components in situ using super-resolution cryo-correlative light and electron microscopy. |
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| المؤلفون: | Last MGF; Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands., Voortman LM; Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands., Sharp TH; Department of Cell and Chemical Biology, Leiden University Medical Centre, Leiden, The Netherlands; School of Biochemistry, University of Bristol, Bristol, United Kingdom. Electronic address: t.sharp@bristol.ac.uk. |
| المصدر: | Methods in cell biology [Methods Cell Biol] 2024; Vol. 187, pp. 223-248. Date of Electronic Publication: 2024 Mar 07. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 0373334 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 0091-679X (Print) Linking ISSN: 0091679X NLM ISO Abbreviation: Methods Cell Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: New York, Academic Press |
| مواضيع طبية MeSH: | Cryoelectron Microscopy*/methods, Humans ; Single Molecule Imaging/methods ; Electron Microscope Tomography/methods ; Software ; Image Processing, Computer-Assisted/methods ; Vimentin/metabolism ; Animals |
| مستخلص: | Super-resolution cryo-correlative light and electron microscopy (SRcryoCLEM) is emerging as a powerful method to enable targeted in situ structural studies of biological samples. By combining the high specificity and localization accuracy of single-molecule localization microscopy (cryoSMLM) with the high resolution of cryo-electron tomography (cryoET), this method enables accurately targeted data acquisition and the observation and identification of biomolecules within their natural cellular context. Despite its potential, the adaptation of SRcryoCLEM has been hindered by the need for specialized equipment and expertise. In this chapter, we outline a workflow for cryoSMLM and cryoET-based SRcryoCLEM, and we demonstrate that, given the right tools, it is possible to incorporate cryoSMLM into an established cryoET workflow. Using Vimentin as an exemplary target of interest, we demonstrate all stages of an SRcryoCLEM experiment: performing cryoSMLM, targeting cryoET acquisition based on single-molecule localization maps, and correlation of cryoSMLM and cryoET datasets using scNodes, a software package dedicated to SRcryoCLEM. By showing how SRcryoCLEM enables the imaging of specific intracellular components in situ, we hope to facilitate adoption of the technique within the field of cryoEM. (Copyright © 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.) |
| فهرسة مساهمة: | Keywords: CLEM; Correlative light-electron microscopy; Cryo-electron tomography; SRcryoCLEM; Super-resolution light microscopy; cryoEM |
| المشرفين على المادة: | 0 (Vimentin) |
| تواريخ الأحداث: | Date Created: 20240505 Date Completed: 20240505 Latest Revision: 20240505 |
| رمز التحديث: | 20240506 |
| DOI: | 10.1016/bs.mcb.2024.02.027 |
| PMID: | 38705626 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0091-679X |
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| DOI: | 10.1016/bs.mcb.2024.02.027 |