Quantitative Super-Resolution Imaging of ER-Phagy Initiation in Cells
Balakrishnan A, Glogger M, Heilemann M (2024)
Publication Type: Journal article
Publication year: 2024
Journal
Book Volume: 2845
Pages Range: 127-140
DOI: 10.1007/978-1-0716-4067-8_10
Abstract
Selective autophagy of the endoplasmic reticulum (ER-phagy) is a mechanism that is necessary for degrading damaged ER components and preventing cells from experiencing ER stress. Various ER-phagy receptors orchestrate this process by building protein assemblies with dedicated functions. In order to understand the molecular building principles of ER-phagy, it is important to reveal the assembly of ER-phagy receptors in a temporal and functional context. However, direct visualization is hampered by the diffraction limit in light microscopy. Super-resolution microscopy (SRM) can bypass this limitation and resolve single proteins and nanoscale protein clusters in cells. In particular, DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful technology that can resolve individual protein clusters in cells and provide information on their molecular composition. Here, we report a step-by-step protocol on how to utilize DNA-PAINT to perform super-resolution imaging of ER-phagy receptors in fixed cells. In addition, we provide a detailed explanation of image generation, cluster analysis, and molecular quantification.
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Germany (DE)How to cite
APA:
Balakrishnan, A., Glogger, M., & Heilemann, M. (2024). Quantitative Super-Resolution Imaging of ER-Phagy Initiation in Cells. Methods in Molecular Biology, 2845, 127-140. https://doi.org/10.1007/978-1-0716-4067-8_10
MLA:
Balakrishnan, Ashwin, Marius Glogger, and Mike Heilemann. "Quantitative Super-Resolution Imaging of ER-Phagy Initiation in Cells." Methods in Molecular Biology 2845 (2024): 127-140.
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