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Expansion Microscopy of Ciliary Proteins

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Cilia

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2725))

Abstract

Label-retention expansion microscopy (LR-ExM) is a sample preparation technique, which embeds the cells or tissues in a swellable hydrogel and expands the sample so that one can achieve a high resolution with any conventional fluorescence microscopes. Fluorescence loss during polymerization and protein denaturation have been a major limitation of standard expansion microscopy. To minimize fluorescence loss, LR-ExM uses trifunctional anchors, which can survive from polymerization and denaturation, and then introduce fluorophores after expansion. By using LR-ExM, one can study the structure of primary cilia at molecular-scale resolution with a much higher signal-to-noise ratio, compared with previously introduced expansion microscopy methods. In this chapter, we describe a detailed procedure showing how LR-ExM is used to study ciliary proteins.

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Acknowledgments

This work was supported by the U.S. National Institutes of Health (R00 GM126136 to X.S.), and partially supported by the U.S. National Science Foundation (DMS1763272 to S.P.), and a grant from the Simons Foundation (594598 to S.P.).

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Authors and Affiliations

  1. Center for Complex Biological Systems, University of California Irvine, Irvine, CA, USA

    Sohyeon Park & Xiaoyu Shi

  2. Department of Developmental and Cell Biology, University of California Irvine, Irvine, CA, USA

    Xiaoyu Shi

  3. Department of Chemistry, University of California Irvine, Irvine, CA, USA

    Xiaoyu Shi

Authors
  1. Sohyeon Park
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  2. Xiaoyu Shi
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Correspondence to Xiaoyu Shi .

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Editors and Affiliations

  1. Medical Research Council Toxicology Unit, School of Biological Sciences, University of Cambridge, Cambridge, UK

    Vito Mennella

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Park, S., Shi, X. (2024). Expansion Microscopy of Ciliary Proteins. In: Mennella, V. (eds) Cilia. Methods in Molecular Biology, vol 2725. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3507-0_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3507-0_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3506-3

  • Online ISBN: 978-1-0716-3507-0

  • eBook Packages: Springer Protocols

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