Data from: Optogenetic neuromuscular actuation of a miniature electronic biohybrid robot
Data files
Apr 07, 2026 version files 2.68 MB
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Dataset_.zip
2.67 MB
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README.md
6.25 KB
Abstract
Neuronal control of skeletal muscle function is ubiquitous across species for locomotion and doing work. Especially, emergent behaviors of neurons in biohybrid neuromuscular systems can advance bioinspired locomotion research. Although recent studies have demonstrated that chemical or optogenetic stimulation of neurons can control muscular actuation through the neuromuscular junction (NMJ), the correlation between neuronal activities and resulting modulation in the muscle responses is less understood, hindering the engineering of high-level functional biohybrid systems. Here, we develop NMJ-based biohybrid crawling robots with optogenetic mouse motor neurons, skeletal muscles, 3D-printed hydrogel scaffolds, and integrated on-board wireless micro light-emitting diode (μLED)-based optoelectronics. We investigate the coupling of the light stimulation and neuromuscular actuation through power spectral density (PSD) analysis. We verify the modulation of the mechanical functionality of the robot depending on the frequency of the optical stimulation to the neural tissue. We demonstrate continued muscle contraction up to 20 minutes after a 1 minute long pulsed 2 hertz optical stimulation of the neural tissue. Furthermore, the robots were shown to maintain their mechanical functionality for over 2 weeks. This study provides insights into reliable neuronal control with optoelectronics, supporting advancements in neuronal modulation, biohybrid intelligence, and automation.
Dataset DOI: 10.5061/dryad.pk0p2nh21
Description of the data and file structure
This dataset contains data obtained from each experiment. The dataset is needed to generate figures and results for the article titled 'Optogenetic Neuromuscular Actuation of a Miniature Electronic Biohybrid Robot.'
Files and variables
File: Dataset_.zip
Description: These data are needed to reproduce the figures in the article titled 'Optogenetic Neuromuscular Actuation of a Miniature Electronic Biohybrid Robot.'
File 1_name: Fig. 2B.csv, description: Relative quantification (RQ) of NMJ-related genes functioning on development and stabilization of acetylcholine receptors. The file contains primer sequences used to the PCR analysis. Ct value means the threshold cycles for the amplification.
File 2_name: Fig. 2C.csv, description: Analysis of fold change of RNA expression between muscle-only and neuromuscular tissues.
File 3_name: Fig. 2D.csv, description: Biological gene ontology analysis based on differentially expressed gene
File 4_name: Fig. 3C.csv, description: Leg deflections of long and short leg obtained from simulation and experimental recording.
File 5_name: Fig. 3E.csv, description: Comparison of crawling trajectories extracted from simulation and experiment.
File 6_name: Fig. 3G.csv, description: Recorded crawling trajectories in x-y plane during chemical inhibitor treatment
File 7_name: Fig. 3H.csv, description: Corresponding net displacement with increasing concentration of the curare.
File 8_name: Fig. 3I.csv, description: Effect of curare and L-glutamic acid on leg deflections.
File 9_name: Fig. 3J.csv, description: Statistics of contraction forces in the presence of chemicals.
File 10_name: Fig. 4C.csv, description: Crawling trajectory of the two-chamber single-NT crawler over time.
File 11_name: Fig. 4D.csv, description: Synchronized twitching of both legs under stimulation to a neural tissue.
File 12_name: Fig. 4E.csv, description: Observation of long leg twitching under 2 Hz and 4 Hz frequencies of optical stimulation.
File 13_name: Fig. 4F.csv, description: Representative deflection traces obtained from each section in Fig. 4E.
File 14_name: Fig. 4G.csv, description: Power spectral densities (PSDs) of muscle twitching modes with respect to stimulus conditions.
File 15_name: Fig. 4H.csv, description: Long-term measurement of muscle twitching after the 2Hz stimulation.
File 16_name: Fig. 5C.csv, description: Comparison of crawling behaviors occurring in non-optogenetic pristine muscle and NMJ crawlers.
File 17_name: Fig. 5D-ii.csv, description: Optimization of the number of neural cells via measuring autonomous crawling of NMJ crawlers.
File 18_name: Fig. 5E-ii.csv, description: A case study of an optoelectronics-driven NMJ crawler that did not crawl before stimulation showed boosted crawling after stimulation.
File 19_name: Fig. 5F-ii.csv, description: A crawler showing high speed of autonomous crawling before stimulation but reduced its velocity after stimulation.
File 20_name: Fig. 6A.csv, description: A representative example of altering muscular actuation depending on neurostimulation frequency (1–4 Hz).
File 21_name: Fig. 6B.csv, description: Long-term performance of a NMJ crawler was verified by measuring the mechanical responses of the robot for 21 days. The data is obtained co-culture day 6.
File 22_name: Fig. 6C.csv, description: Long-term performance of a NMJ crawler was verified by measuring the mechanical responses of the robot for 21 days. The data is obtained co-culture day 12.
File 23_name: Fig. 6D.csv, description: Measured muscle contraction forces in these long-term measurement experiment in a NMJ biohybrid crawler showing functionality lasting over 2 weeks.
File 24_name: Fig. S5.csv, description: Traces of calcium flux occurring in the NMJ, showing spontaneous neuronal firing and muscle contraction.
File 25_name: Fig. S6.csv, description: Representative NMJ crawlers showing autonomous crawling locomotion.
File 26_name: Fig. S7B.csv, description: FEA simulation of leg deflection in a dual-NT crawler with independent neural signaling frequency.
File 27_name: Fig. S7C.csv, description: FEA simulation of leg deflection in a dual-NT crawler with independent neural signaling with lagging conditions.
File 28_name: Fig. S9.csv, description: Quantitative analysis on leg deflection under 2Hz and 4 Hz opto-stimulations.
File 29_name: Fig. S13.csv, description: Investigation of the time required to modulate the twitching mode.
File 30_name: Fig. S14.csv, description: Crawling traces of a NMJ crawler under single frequency (2Hz and 4Hz) of optical stimulation.
File 31_name: Fig. S15A.csv, description: Twitching modulation via opto-stimulation of a single-NT NMJ robot and analysis of crawling dynamics. Data show the full traces of the recording.
File 32_name: Fig. S15C.csv, description: Twitching modulation via opto-stimulation of a single-NT NMJ robot and analysis of crawling dynamics. Data show the measured forces and standard deviation at each condition.
File 33_name: Fig. S15D.csv, description: Twitching modulation via opto-stimulation of a single-NT NMJ robot and analysis of crawling dynamics. Data show the calculated crawling velocity obtained from the force and twitching frequency.
File 34_name: Fig. S15E.csv, description: Twitching modulation via opto-stimulation of a single-NT NMJ robot and analysis of crawling dynamics. Data show the simulated crawling traces based on the velocity described in the Fig. S15D.
File 35_name: Fig. S16.csv, description: Effect of the muscle thickness and friction coefficient on crawling dynamics.
File 36_name: Fig. S17.csv, description: Different crawling behavior between single or dual-neural-tissue (NT) model.
File 37_name: Fig. S18.csv, description: Power spectral densities obtained from multiple NMJ crawlers which responded to the 2 Hz stimulation.
File 38_name: Fig. S19.csv, description: Long-term measurement of the neuromuscular actuation with 2Hz optical stimulation.