Three-photon head-mounted microscope for imaging deep cortical layers in freely moving rats

We designed a head-mounted three-photon microscope for imaging deep cortical layer neuronal activity in a freely moving rat. Delivery of high-energy excitation pulses at 1,320 nm required both a hollow-core fiber whose transmission properties did not change with fiber movement and dispersion compensation. These developments enabled imaging at >1.1 mm below the cortical surface and stable imaging of layer 5 neuronal activity for >1 h in freely moving rats performing a range of behaviors.

Klioutchnikov, A., Wallace, D.J., Frosz, M.H., Zeltner, R., Sawinski, J., Pawlak, V., Voit, K.M., Russel, P.St.J. and Kerr, J.N.D. (2020). Three-photon head-mounted microscope for imaging deep cortical layers in freely moving rats. Nat. Methods 17, 509-513.
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• Center for Advanced European Studies and Research