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. Author manuscript; available in PMC: 2017 Aug 19.
Published in final edited form as: Nat Med. 2016 Nov 21;23(1):49–59. doi: 10.1038/nm.4233

Figure 3.

Figure 3

Live imaging of neural activity in HIOs+ENS. (a) Live imaging of Ca2+ flux in the neural-crest-derived ENS cells revealed periodic activity. Shown are snapshots from a 20-min time-lapse video (see Supplementary Video 3) of neural activity in HIOs+ENS. Colored arrows point to cells whose pixel intensity was measured over time. Numbers in snapshots correspond to numbered peaks in the graph. The graph measures fluorescence values as ΔF/F0, (ΔF = Ft – F0), where Ft is observed fluorescence at time t and F0 is fluorescence at t = 0. The color of the line corresponds to the same colored arrowhead in the snapshots. Scale bars, 50 μm. (b) Live imaging of Ca2+ flux in HIOs+ENS grown in vitro before (left) and after KCl treatment, which induced a rapid calcium efflux in the ENS cells (see Supplementary Video 4). Data are representative of two independent experiments. Scale bars, 100 μm. (c) Live imaging of Ca2+ flux in HIOs+ENS grown in vivo shows calcium transients in nerve bundles before and after KCl treatment, which induces a broad calcium efflux and contraction of the tissue (see Supplementary Videos 5 and 6). In vivo data are representative of two independent experiments with n = 3 organoids engrafted into individual mice per experiment. Scale bars, 100 μm (left) and 500 μm (right).