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Expansion Microscopy of Plant Cells (PlantExM).

Timothy J Hawkins1, Joanne L Robson2, Bethany Cole2

  • 1Department of Biosciences, Durham University, Durham, UK. t.j.hawkins@durham.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2023
PubMed
Summary

Expansion microscopy (ExM) enables super-resolution imaging of plant cells without specialized hardware. This method expands samples, resolving fine structures like the cytoskeleton in unprecedented detail.

Keywords:
Cell divisionClearingExpansion microscopyFluorescenceLightsheetRootsSuper-resolutionTobacco bright yellow 2 (BY2)

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Area of Science:

  • Biotechnology
  • Microscopy
  • Plant Biology

Background:

  • Expansion microscopy (ExM) offers super-resolution imaging by physically and optically magnifying samples.
  • Conventional ExM is adapted for plants, requiring specific protocols for cell wall digestion.

Purpose of the Study:

  • To develop and optimize an ExM method (PlantExM) for high-resolution plant cytoskeleton imaging.
  • To overcome limitations in plant super-resolution microscopy, such as cell size and light scattering.

Main Methods:

  • Plant samples are fixed, stained, and embedded in a swellable gel matrix.
  • Sample components are digested, and the gel is expanded in water to increase resolution.
  • PlantExM is optimized for cytoskeleton visualization, compatible with optical super-resolution techniques.

Main Results:

  • PlantExM successfully achieves super-resolution imaging of the plant cytoskeleton.
  • The method enables detailed visualization of subcellular structures within plant tissues at depth.
  • This technique overcomes previous limitations posed by plant cell walls and tissue optical properties.

Conclusions:

  • PlantExM provides a powerful tool for high-resolution plant cytoskeleton imaging.
  • This advancement opens new avenues for studying plant cell biology at the nanoscale.
  • The method facilitates unprecedented detail in plant super-resolution microscopy.