Abstract
Expansion microscopy is an imaging method based on isotropic physical expansion of biological samples, which improves optical resolution and allows imaging of subresolutional cellular components by conventional microscopes. Centrioles are small microtubule-based cylindrical structures that build centrosomes and cilia, two organelles essential for vertebrates. Due to a centriole’s small size, electron microscopy has traditionally been used to study centriole length and ultrastructural features. Recently, expansion microscopy has been successfully used as an affordable and accessible alternative to electron microscopy in the analysis of centriole and cilia length and structural features. Here, we describe an expansion microscopy approach for the analysis of centrioles and cilia in large populations of mammalian adherent and nonadherent cells and multiciliated cultures.
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Acknowledgments
We thank members of LPDS and Dr. Valentin Magidson for critical reading of the manuscript, and Dr. Catherine Sullenberger for acquiring images used in Fig. 3d. This work was supported by the Intramural Research Program of the National Institutes of Health (NIH), National Cancer Institute to J.L.
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Kong, D., Loncarek, J. (2021). Analyzing Centrioles and Cilia by Expansion Microscopy. In: Coutts, A.S., Weston, L. (eds) Cell Cycle Oscillators . Methods in Molecular Biology, vol 2329. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1538-6_18
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DOI: https://doi.org/10.1007/978-1-0716-1538-6_18
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