Fig. 4: GraMOS enables functional phenotyping and promotes maturation of G-interfaced hiPSC-derived neurons. | Nature Communications

Fig. 4: GraMOS enables functional phenotyping and promotes maturation of G-interfaced hiPSC-derived neurons.

From: Non-genetic neuromodulation with graphene optoelectronic actuators for disease models, stem cell maturation, and biohybrid robotics

Fig. 4

a Scheme for GraMOS-empowered all-optical studies using single-cell single-pulse light. b Videoframes of G-interfaced Fluo-4-labeled neurons at rest and following activation of a central neuron by a single light pulse (561 nm, 5.2 mW/mm2). c Calcium transients across WTAD and M233L networks triggered by a single light pulse. d Ratio of GraMOS-activated neurons in WTAD and M233L models (n = 9 and 18 G-coverslips, respectively; **P = 0.0098, one-way ANOVA). e Calcium transient amplitudes in G-interfaced WTAD and M233L neurons (n = 251 and 678, respectively; ***P = 0.0004, one-way ANOVA) activated by a single light pulse. Box plots in (d) and (e): center line = median, red squares = mean, box = upper/lower quartiles, whiskers = 5th–95th percentiles, and individual points = circles. f Spearman correlation matrix for GraMOS-initiated neuronal activity in G-interfaced WTAD and M233L neurons. g Spatial maps showing the number of spikes in GraMOS-activated WTAD and M233L neuronal networks. h Spatio-temporal maps depicting the activation times (in seconds) after a light pulse in WTAD and M233L neurons (****P = 1.0063 × 10-235, two-sided Wilcoxon rank sum). i Histogram of activation times for WTAD and M233L models. j Scheme for GraMOS-driven maturation of G-interfaced hiPSC-neurons. k Ratio of GraMOS-activated WTAD neurons at different timepoints during light training (n = 47, 16, 9, 20, 15 for Weeks 0, 2, 3, 4, 6) (*P = 0.0422 (Weeks 0 vs 3); ***P = 0.0006 (Weeks 2 vs 4); ****P = 8.119 × 10−7 (Weeks 0 vs 4); P for Week 6 vs Week 0–6.6059 × 10−18, Week 2–8.2329 × 10−7, Week 3–1.3661 × 10−7, Week 4–0.0011; Two-way ANOVA). l Amplitudes of GraMOS-triggered calcium transients in WTAD neurons at different timepoints during light training (n = 542, 202, 245, 364, 675 neurons for Weeks 0, 2, 3, 4, 6) (****P = 7.0376 × 10−7, (Weeks 0 vs 3); ***P = 3.1209 × 10−4 (Weeks 0 vs 2); ****P = 5.154 × 10−5 (Weeks 0 vs 4); P = 2.5673 × 10−8 (Weeks 0 vs 6); Two-way ANOVA). Box plots in (k) and (l): center line = median, red squares = mean, box = upper/lower quartiles, whiskers = 5th–95th percentiles, and individual points = circles. m Spearman correlation matrix for GraMOS-initiated neuronal activity in WTAD models at different timepoints during light training. A.U. arbitrary units.

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