دورية أكاديمية
Enhanced expansion microscopy to measure nanoscale structural and biochemical remodeling in single cells.
| العنوان: | Enhanced expansion microscopy to measure nanoscale structural and biochemical remodeling in single cells. |
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| المؤلفون: | Sheard TMD; Faculty of Biological Sciences, School of Biomedical Sciences, University of Leeds, Leeds, United Kingdom., Jayasinghe I; Faculty of Biological Sciences, School of Biomedical Sciences, University of Leeds, Leeds, United Kingdom; Department of Molecular Biology & Biotechnology, Faculty of Science, The University of Sheffield, Sheffield, United Kingdom. Electronic address: i.jayasinghe@sheffield.ac.uk. |
| المصدر: | Methods in cell biology [Methods Cell Biol] 2021; Vol. 161, pp. 147-180. Date of Electronic Publication: 2020 Jun 10. |
| نوع المنشور: | Journal Article |
| 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: | Microscopy, Confocal* |
| مستخلص: | Resolution is a key feature in microscopy which allows the visualization of the fine structure of cells. Much of the life processes within these cells depend on the three-dimensional (3D) complexity of these structures. Optical super-resolution microscopies are currently the preferred choice of molecular and cell biologists who seek to visualize the organization of specific protein species at the nanometer scale. Traditional super-resolution microscopy techniques have often been limited by sample thickness, axial resolution, specialist optical instrumentation and computationally-demanding software for assembling the images. In this chapter we detail the protocol, "enhanced expansion microscopy" (EExM), which combines X10 expansion microscopy with Airyscan confocal microscopy. EExM enables 15nm lateral (and 35nm axial) resolution, and is a relatively cheap, accessible option allowing single protein resolution for the non-specialist optical microscopists. We illustrate how EExM has been utilized for mapping the 3D topology of intracellular protein arrays at sample depths which are not always compatible with some of the traditional super-resolution techniques. We demonstrate that antibody markers can recognize and map post-translational modifications of individual proteins in addition to their 3D positions. Finally, we discuss the current uncertainties and validations in EExM which include the isotropy in gel expansion and assessment of the expansion factor (of resolution improvement). (© 2021 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Airyscan; Biochemical features; Enhanced expansion microscopy; Nanodomains; Nanoscale; Remodeling; Single cells; Super-resolution; X10 expansion microscopy |
| تواريخ الأحداث: | Date Created: 20210122 Date Completed: 20211125 Latest Revision: 20211125 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/bs.mcb.2020.04.019 |
| PMID: | 33478687 |
| قاعدة البيانات: | MEDLINE |
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