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Expansion Microscopy of Plant Cells (PlantExM)

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The Plant Cytoskeleton

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2604))

Abstract

Expansion microscopy (ExM) achieves super-resolution imaging without the need for sophisticated super-resolution microscopy hardware through a combination of physical and optical magnification. Samples are fixed, stained, and embedded in a swellable gel. Following cross-linking of fluorophores to the gel matrix, the components of the sample are digested away and the gel expanded in water. Labeled objects which are too close to be resolved by diffraction-limited microscopy are moved far enough apart that these can now be resolved as individual objects on a standard confocal. Originally developed for animal cells and tissues, ExM for plants requires the additional consideration of cell wall digestion. Super-resolution can be limited in plants due to the size of cells, light scattering of tissues, and variations in refractive index. By removing the components which cause these limitations, ExM opens up the possibility of super-resolution at depth within plant tissues for the first time. Here we describe our method for PlantExM which is optimized for cytoskeleton resolution, which, when also coupled with compatible optical super-resolution technologies, can produce images of the plant cytoskeleton in unprecedented detail.

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Author information

Authors and Affiliations

  1. Department of Biosciences, Durham University, Durham, UK

    Timothy J. Hawkins, Joanne L. Robson, Bethany Cole & Simon J. Bush

Authors
  1. Timothy J. Hawkins
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  2. Joanne L. Robson
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  3. Bethany Cole
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  4. Simon J. Bush
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Corresponding author

Correspondence to Timothy J. Hawkins .

Editor information

Editors and Affiliations

  1. Department of Biosciences, University of Durham, Durham, UK

    Patrick J. Hussey

  2. College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, China

    Pengwei Wang

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Hawkins, T.J., Robson, J.L., Cole, B., Bush, S.J. (2023). Expansion Microscopy of Plant Cells (PlantExM). In: Hussey, P.J., Wang, P. (eds) The Plant Cytoskeleton. Methods in Molecular Biology, vol 2604. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2867-6_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2867-6_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2866-9

  • Online ISBN: 978-1-0716-2867-6

  • eBook Packages: Springer Protocols

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