Fig. 2

Setup of the saphenous nerve imaging experiment. a After deep anesthesia, the mouse is placed on its back, the skin of its left inner tight is removed and the saphenous nerve (N) is placed on a plastic strip (S). Two stimulation microelectrodes (1 and 2) are inserted on both sides of the nerve on the plastic strip and one hook-shaped recording electrode (3) is holding the nerve on the opposite side. b The ground electrode (4) is inserted in the mouse tail and the negative electrode (5) in the groin area. c A glass coverslip (Co) is then placed on the nerve soaked in PBS and the mouse is then placed under the two-photon microscope immersion lens 20X (L) in water. d Schematic view of the imaging setup. e Schematic representation of the nerve stimulation pattern used to induce APs. The generation of APs was verified using the recording electrode. Negative electrode was used to correct for background signal