Fig. 8

In vivo Ca²⁺ imaging of spinal dorsal horn neurons. a A scheme of experimental setup. Two-photon imaging of SDH was performed through custom built implanted imaging chamber and electrode was inserted into one side of ACC after a small craniotomy. Pinching and brushing were applied to the contralateral hindpaw. b–c Representative image showing the Ca²⁺ responses before and after pinching/brushing and ACC stimuli. Raw fluorescent intensity was pseudo-colored and region of interests were numbered. Changes of intensity were plotted under the images and corresponding time points to images were indicated by open arrow head (Pinching/Brushing) and closed arrow head (basal). Roi 1, 2, 6 in b and Roi 1, 2, 4 in c were identified neurons with potentiated responses to ACC stimulation; Roi 3 or Roi 4, 5 in b were neurons with alleviated or no changed responses. Roi 3, 4, 6–8 in c have no responses to brushing. d Maximal Ca²⁺ response to brushing and pinching in each cells indicated as ΔF_t/F₀ were plotted at pre-ACC and post-ACC stimulation. Data were separately plotted on the basis of the effect of ACC stimulation (increase or decrease). e Summarized results of identified cells before and after pinching/brushing and ACC stimuli