In vivo superresolution imaging of neuronal structure in the mouse brain

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

*Corresponding author for this work

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Objective: this study proposes and evaluates a technique for in vivo deep-tissue superresolution imaging in the light-scattering mouse brain at up to a 3.5 Hz 2-D imaging rate with a 21×21 μm2 field of view. Methods: we combine the deep-tissue penetration and high imaging speed of resonant laser scanning two-photon (2P) microscopy with the superresolution ability of patterned excitation microscopy. Using high-frequency intensity modulation of the scanned two-photon excitation beam, we generate patterned illumination at the imaging plane. Using the principles of structured illumination, the high-frequency components in the collected images are then used to reconstruct images with an approximate twofold increase in optical resolution. Results: using our technique, resonant 2P superresolution patterned excitation reconstruction microscopy, we demonstrate our ability to investigate nanoscopic neuronal architecture in the cerebral cortex of the mouse brain at a depth of 120 μm in vivo and 210 μm ex vivo with a resolution of 119 nm. This technique optimizes the combination of speed and depth for improved in vivo imaging in the rodent neocortex. Conclusion: this study demonstrates a potentially useful technique for superresolution in vivo investigations in the rodent brain in deep tissue, creating a platform for investigating nanoscopic neuronal dynamics. Significance: this technique optimizes the combination of speed and depth for improved superresolution in vivo imaging in the rodent neocortex.

Original languageEnglish (US)
Pages (from-to)232-238
Number of pages7
JournalIEEE Transactions on Biomedical Engineering
Volume65
Issue number1
DOIs
StatePublished - Jan 2018

Keywords

  • Neuron
  • Structured illumination
  • Super-resolution imaging
  • Two-photon microscopy

ASJC Scopus subject areas

  • Biomedical Engineering

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