Fig. 1: Real-time luminescence recording for live measurement of clock gene dynamics.

a Schematic representation of experimental setup for the acquisition of bioluminescence signal from cell integrated microfluidic chip; microfluidic setup is coupled with a microscope and liquid handling system. b Schematic representation of the liquid handling system connected to the microfluidic device, composed of two independent culture chambers (two inlets and one outlet for each). Scale bar 2.5 mm. c Time-lapse of bioluminescence images of a representative experiment, captured every 30 min, of mouse Per2::Luc fibroblasts, showing circadian rhythms of luminescence for 2 days. Scale bar 1 mm. d Baseline-subtracted Per2::Luc bioluminescence patterns acquired for 4 days from a microfluidic chamber after dexamethasone shock. Data are represented as mean ± s.d., N = 9 for each condition. e Temporal profiles of clock gene expressions, BMAL1 and PER2, in human fibroblasts integrated in the microfluidic platform, after synchronization (t = 0) with dexamethasone. mRNA level of clock genes was measured by qPCR and normalized to GADPH expression. Data are represented as mean ± s.d., N = 3 for each group. f Top, Schematic representation of the two different protocols of medium delivery imposed by automatic medium change. Bottom, Per2::Luc 6-day bioluminescence signal intensity in the 16-by-16 pixel ROIs, shown in the scheme in h (top), under the two medium change conditions, after baseline subtraction using LumiCycle Analysis program (Actimetrics). Solid line: mean signal intensity, patch: area delimited by mean ± s.d. of the 48 ROIs indicated in h. g Heatmaps of mean Per2::Luc 6-day bioluminescence signal intensity in the 16-by-16 pixel ROIs, shown in the schemes on the left, under the two medium change conditions shown in f. Colorbar indicates the bioluminescence signal intensity after image pre-processing in arbitrary units and applies to all four heatmaps. Scale bar 1 mm. h Kuramoto order parameter K, calculated for the time intervals between time 0.5 d and the time shown on the x axis, indicating the level of synchrony between the 16-by-16 pixel ROIs, shown in the scheme on top (K = 1 means complete synchrony). The horizontal gray line highlights a value of 0.7. Source data are available as a Source Data file. Scale bar 1 mm.