Direct wavefront sensing enables functional imaging of infragranular axons and spines

Nat Methods. 2019 Jul;16(7):615-618. doi: 10.1038/s41592-019-0434-7. Epub 2019 Jun 17.


We advance two-photon microscopy for near-diffraction-limited imaging up to 850 µm below the pia in awake mice. Our approach combines direct wavefront sensing of light from a guidestar (formed by descanned fluorescence from Cy5.5-conjugated dextran in brain microvessels) with adaptive optics to compensate for tissue-induced aberrations in the wavefront. We achieve high signal-to-noise ratios in recordings of glutamate release from thalamocortical axons and calcium transients in spines of layer 5b basal dendrites during active tactile sensing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Axons / metabolism*
  • Calcium / metabolism
  • Dendritic Spines / metabolism*
  • Glutamic Acid / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microscopy
  • Signal-To-Noise Ratio


  • Glutamic Acid
  • Calcium