In vivo fluorescence microscopy of neuronal activity in three dimensions by use of voltage-sensitive dyes.
We report in vivo imaging of neuronal electrical activity from superficial layers of the mouse barrel cortex. The measurements have approximately 16-microm spatial and 3-ms temporal resolution and reach depths of 150 microm below the cortical surface. The depth-dependent differential-fluorescence optical sections of activity are consistent with known cortical architecture and represent an important step toward in vivo measurement of functioning complex neural networks. Our observations employ a custom gradient-index lens probe and voltage-sensitive dye fluorescence; the use of epi-illumination rather than dark-field illumination provides the dramatic signal-to-noise improvement necessary for fast three-dimensional imaging.
Fisher
Jonathan A N
JA
Department of Physics and Astronomy, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6396, USA. aafisher@sas.upenn.edu
Civillico
Eugene F
EF
Contreras
Diego
D
Yodh
Arjun G
AG
D000818:Animals; D004396:Coloring Agents; D021621:Imaging, Three-Dimensional; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons; D013003:Somatosensory Cortex / Q000166:cytology / Q000502:physiology*; D013343:Styrenes
D004396:Coloring Agents; C096453:RH 795; D013343:Styrenes