Related to Fig. 6. Optical manipulation of dorsal raphe nucleus (a), inferior olive (b) and nucleus MLF (c) during volumetric functional imaging, using a transgenic line Tg(elavl3:jRGECO1b) x Tg(elavl3:CoChR-eGFP), as in Fig. 6. Targeted cells are indicated by yellow dots and locations of example cell responses by open circles. Shown are four maximum intensity projections, each covering a different 50 µm z-interval of the full image volume (left). The color code indicates the weighted activity change relative to control manipulations, averaged across 12 trials for targets in the DRN, 10 trials for the IO, and 6 trials for the nMLF. Responses are weighted relative to manipulations targeted to points outside the brain to de-emphasize activity responses that are not well conserved across trials and that are shared between manipulation inside and outside the brain. Control manipulations were targeted to positions outside the brain. ΔF/F time traces of the indicated positions (right) show the activity profile of selected single cells (orange, magenta, blue, green) located in the cerebellar cortex, dorsal raphe nucleus (DRN), dorsal hindbrain (DHB) and inferior olive. Traces are shown for all trials (thin colored lines) and controls (thin black lines; for these controls the two-photon laser was targeted just outside the fish’s head), as well as for their respective mean (thick lines).he experiment shown in this figure was repeated independently three times with similar results. Scale bar, 50 µm.