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Chapter 17 Correlative Light and Electron Microscopy
Abstract
Autophagy is an intracellular degradative pathway that is essential for cellular homeostasis. Efficient autophagy ultimately relies on the ability of the cell to form autophagosomes, and the efficiency of lysosomal enzymes and lipid hydrolases contained within the autolysosome to degrade sequestered cytosolic material and organelles and recycle these nutrients back to the cytosol. Several assays and techniques to monitor autophagy are available, and these can be quantitative or qualitative, biochemical or morphological. Here we describe a method for monitoring the autophagic process that is based on morphology and the application of both light and electron microscopy, called correlative light and electron microscopy, or CLEM. CLEM provides an advance over either technique (light or electron microscopy) alone and can be performed on any cell or tissue sample, which can be grown or mounted on a gridded coverslip or support compatible with light microscopy. CLEM gives a broad low magnification overview of the cell, allowing an assessment of both spatial and temporal events, as well as providing high-resolution information about individual autophagosomes or single compartments.
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“Finally, a recent advent of CRISPR-Cas9 gene-editing technologies provides a promising alternative to overcome potential pitfalls of GFP-LC3 overexpression-the generation of knockin cell lines, in which the coding sequence of GFP is added in frame with the 5' sequence (encoding the Nterminal part) of endogenous LC3 [122,407].”
From Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1 (2021) · cites this paper as [122]
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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).
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Self-citationGuidelines for the use and interpretation of assays for monitoring autophagy
Klionsky, D. J., Abdalla, F. C., Abeliovich, H., Abraham, R. T., Acevedo-Arozena, A., Adeli, K., Agholme, L., Agnello, M., Agostinis, P., Aguirre-Ghiso, J. A., et al. (2012) · Autophagy · doi:10.4161/auto.19496 · Cited by 3,996
Self-citationGuidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1
Klionsky, D. J., Abdel-Aziz, A. K., Abdelfatah, S., Abdellatif, M., Abdoli, A., Abel, S., Abeliovich, H., Abildgaard, M. H., Abudu, Y. P., Acevedo-Arozena, A., et al. (2021) · Autophagy · doi:10.1080/15548627.2020.1797280 · Cited by 2,897
Self-citationLC3-Associated Phagocytosis (LAP): Connections with Host Autophagy
Lai, S.-C., & Devenish, R. J. (2012) · Cells · doi:10.3390/cells1030396 · Cited by 116
Metadata
- Authors
- Razi, M., & Tooze, S. A.
- Venue
- Methods in Enzymology
- Publisher
- Elsevier
- Published
- 2009
- Pages
- 261-275
- ISSN
- 0076-6879
- Cited by
- 46
- References
- 0
- Language
- en
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