4 years ago

Imaging neuronal structure dynamics using two-photon super-resolution patterned excitation reconstruction (SuPER) microscopy

Siyu Chen, Biqin Dong, Ben E. Urban, Yevgenia Kozorovitskiy, Lei Xiao, Hao F. Zhang
Visualizing fine neuronal structures deep inside strong 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 three-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.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jbio.201700171

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