Aberration-free three-dimensional multiphoton imaging of neuronal activity at kHz rates.
Multiphoton microscopy is a powerful tool in neuroscience, promising to deliver important data on the spatiotemporal activity within individual neurons as well as in networks of neurons. A major limitation of current technologies is the relatively slow scan rates along the z direction compared to the kHz rates obtainable in the x and y directions. Here, we describe a custom-built microscope system based on an architecture that allows kHz scan rates over hundreds of microns in all three dimensions without introducing aberration. We further demonstrate how this high-speed 3D multiphoton imaging system can be used to study neuronal activity at millisecond resolution at the subcellular as well as the population level.
Department of Engineering Science, University of Oxford, Parks Road, Oxford OX1 3PJ, United Kingdom.
Edward J
EJ
Botcherby
Christopher W
CW
Smith
Michael M
MM
Kohl
Delphine
D
Débarre
Martin J
MJ
Booth
Rimas
R
Juškaitis
Ole
O
Paulsen
Tony
T
Wilson
D000818:Animals; D002540:Cerebral Cortex / Q000166:cytology; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D009474:Neurons / Q000502:physiology*; D015203:Reproducibility of Results