Expression of OLA-1::GFP and ZYG-8::GFP in head region were observed with BX53 upright microscope (Olympus, Tokyo, Japan). OLA-1::GFP expression in the whole body was observed with LSM880 confocal microscope (Zeiss).
Calcium imaging was performed as described elsewhere [37 (link),41 (link)]. Briefly, a single adult animal that expressed genetically encoded calcium indicator GCaMP3 [59 (link)] and/or XCaMP-R [60 (link)] was placed on a 10% agar pad on a cover slip with 0.1 μm polystyrene beads (Polysciences, Warrington, PA, USA) and covered by another cover slip for immobilization [92 (link)]. The immobilized animals were placed on a Peltier-based temperature controller (Tokai Hit, Fujinomiya, Japan) on a stage of BX61WI microscope (Olympus, Tokyo, Japan). The red and green fluorescence was separated by the Dual-View optics system (Molecular Devices, Sunnyvale, CA, USA), and the images were captured by an EM-CCD camera C9100-13 ImageEM (Hamamatsu Photonics, Japan) at 1 frame per second. Excitation pulses were generated by SPECTRA light engine (Lumencor, Beaverton, OR, USA). The fluorescence intensities were measured by the MetaMorph imaging system (Molecular Devices).
Calcium imaging was performed as described elsewhere [37 (link),41 (link)]. Briefly, a single adult animal that expressed genetically encoded calcium indicator GCaMP3 [59 (link)] and/or XCaMP-R [60 (link)] was placed on a 10% agar pad on a cover slip with 0.1 μm polystyrene beads (Polysciences, Warrington, PA, USA) and covered by another cover slip for immobilization [92 (link)]. The immobilized animals were placed on a Peltier-based temperature controller (Tokai Hit, Fujinomiya, Japan) on a stage of BX61WI microscope (Olympus, Tokyo, Japan). The red and green fluorescence was separated by the Dual-View optics system (Molecular Devices, Sunnyvale, CA, USA), and the images were captured by an EM-CCD camera C9100-13 ImageEM (Hamamatsu Photonics, Japan) at 1 frame per second. Excitation pulses were generated by SPECTRA light engine (Lumencor, Beaverton, OR, USA). The fluorescence intensities were measured by the MetaMorph imaging system (Molecular Devices).