Imaging neuronal structure dynamics using 2-photon super-resolution patterned excitation reconstruction microscopy

Ben E. Urban, Lei Xiao, Biqin Dong, Siyu Chen, Yevgenia Kozorovitskiy*, Hao F. Zhang

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Visualizing fine neuronal structures deep inside strongly light-scattering brain tissue remains a challenge in neuroscience. Recent nanoscopy techniques have reached the necessary resolution but often suffer from limited imaging depth, long imaging time or high light fluence requirements. Here, we present two-photon super-resolution patterned excitation reconstruction (2P-SuPER) microscopy for 3-dimensional imaging of dendritic spine dynamics at a maximum demonstrated imaging depth of 130 μm in living brain tissue with approximately 100 nm spatial resolution. We confirmed 2P-SuPER resolution using fluorescence nanoparticle and quantum dot phantoms and imaged spiny neurons in acute brain slices. We induced hippocampal plasticity and showed that 2P-SuPER can resolve increases in dendritic spine head sizes on CA1 pyramidal neurons following theta-burst stimulation of Schaffer collateral axons. 2P-SuPER further revealed nanoscopic increases in dendritic spine neck widths, a feature of synaptic plasticity that has not been thoroughly investigated due to the combined limit of resolution and penetration depth in existing imaging technologies.

Original languageEnglish (US)
Article numbere201700171
JournalJournal of Biophotonics
Issue number3
StatePublished - Mar 2018


  • dendritic spine
  • neuron
  • nonlinear optics
  • super-resolution microscopy

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Engineering(all)
  • Physics and Astronomy(all)


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