Imaging nanoscale nuclear structures with expansion microscopy

Emma Faulkner, Jeremy Pike, Ruth Densham, Evelyn Garlick, Steven Thomas, Robert Neely*, Jo Morris*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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Commonly applied super-resolution light microscopies have provided insight into subcellular processes at the nanoscale. However, imaging depth, speed, throughput and cost remain significant challenges, limiting the numbers of three-dimensional (3D) nanoscale processes that can be investigated and the number of laboratories able to undertake such analysis. Expansion microscopy (ExM) solves many of these limitations, but its application to imaging nuclear processes has been constrained by concerns of unequal nuclear expansion. Here, we demonstrate the conditions for isotropic expansion of the nucleus at a resolution equal to or better than 120-130 nm (pre-expansion). Using the DNA damage response proteins BRCA1, 53BP1 (also known as TP53BP1) and RAD51 as exemplars, we quantitatively describe the 3D nanoscale organisation of over 50,000 DNA damage response structures. We demonstrate the ability to assess chromatin-regulated events and show the simultaneous assessment of four elements. This study thus demonstrates how ExM can contribute to the investigation of nanoscale nuclear processes.

Original languageEnglish
Article numberjcs259009
Number of pages11
JournalJournal of Cell Science
Issue number14
Publication statusPublished - 19 Jul 2022

Bibliographical note

Funding Information:
The authors acknowledge the help of Dee Kavanagh (COMPARE), Iain Styles (University of Birmingham), Krystian Ubych (University of Birmingham), Alexandra Walker (University of Birmingham) and James Beesley (University of Birmingham) for advice in development of the project. We acknowledge Engineering and Physical Sciences Research Council (EPSRC; UK Research and Innovation), grant numbers EP/L016346/1 (EPSRC Centre for Doctoral Training in Physical Sciences for Health, Sci-Phy-4-Health) and EP/N020901/1 for funding. J.R.M. and R.M.D. are supported by the University of Birmingham. S.G.T. received funding from the British Heart Foundation (NH/18/3/33913) and COMPARE. Open access funding provided by University of Birmingham. Deposited in PMC for immediate release.

Publisher Copyright:
© 2022. Published by The Company of Biologists Ltd.


  • DNA
  • DNA damage response
  • Expansion microscopy
  • Fluorescence
  • Microscopy
  • Super-resolution
  • Exploring the nucleus
  • 53BP1
  • DNA damage
  • BRCA1
  • Nanoscale
  • RAD51
  • Imaging
  • Tools in cell biology
  • Chromatin
  • Microscopy/methods
  • Cell Nucleus


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