High-throughput rhythmic regulation of cardiomyocytes by integrated electrical stimulation and video-based automated analysis biosensing platform.

In cardiac tissue engineering, electric stimulation is an efficient approach to improve the formation of cardiac tissue from individual cardiomyocyte. The regulation conditions of electric stimulation should be screened in an efficient way. However, the lack of high-throughput and large-scale assessment platforms limited the effectively screen the regulation conditions. Here, we develop a high-throughput integrated electrical stimulation system to rhythmically regulate the cardiomyocytes in situ. The state of regulated cardiomyocytes is characterized by a video-based automated biosensing system to analyze the beating of cardiomyocytes. Electrical stimulation conditions are optimized to regulate the cardiomyocyte state in vitro to replace the complex bioactive molecules and materials. By the video analysis, the accurate beating rate and regularity of cardiomyocyte can be determined. The results show that electrical stimulation frequency is a significant factor to regulate the cardiomyocyte beating. The electrical stimulation with a frequency of 3 Hz can effectively regulate the primary rat cardiomyocytes with normal rhythm. This high-throughput electrical stimulation and a video-based automated biosensing system will be a promising and powerful tool to effectively optimize the regulation conditions of cardiomyocyte in vitro, and possess broad application prospects in cardiac tissue engineering and pharmaceutical industry.

A universal, multimodal cell-based biosensing platform for optimal intracellular action potential recording.

Intracellular recording of action potentials is an essential mean for studying disease mechanisms, and for electrophysiological studies, particularly in excitable cells as cardiomyocytes or neurons. Current strategies to obtain intracellular recordings include three-dimensional (3D) nanoelectrodes that can effectively penetrate the cell membrane and achieve high-quality intracellular recordings in a minimally invasive manner, or transient electroporation of the membrane that can yield temporary intracellular access. However, the former strategy requires a complicated and costly fabrication process, and the latter strategy suffers from high dependency on the method of application of electroporation, yielding inconsistent, suboptimal recordings. These factors hinder the high throughput use of these strategies in electrophysiological studies. In this work, we propose an advanced cell-based biosensing platform that relies on electroporation to produce consistent, high-quality intracellular recordings. The suggested universal system can be integrated with any electrode array, and it enables tunable electroporation with controllable pulse parameters, while the recorded potentials can be analyzed in real time to provide instantaneous feedback on the electroporation effectiveness. This integrated system enables the user to perform electroporation, record and assess the obtained signals in a facile manner, to ultimately achieve stable, reliable, intracellular recording. Moreover, the proposed platform relies on microelectrode arrays which are suited for large-scale production, and additional modules that are low-cost. Using this platform, we demonstrate the tuning of electroporation pulse width, pulse number, and amplitude, to achieve effective electroporation and high-quality intracellular recordings. This integrated platform has the potential to enable larger scale, repeatable, convenient, and low-cost electrophysiological studies.

Porous Polyethylene Terephthalate Nanotemplate Electrodes for Sensitive Intracellular Recording of Action Potentials.

New strategies for intracellular electrophysiology break the spatiotemporal limitation of the action potential and lead a notable advance in the investigation of electrically excitable cells and their network. Although successful applications of intracellular recording have been achieved by 3D micro/nanodevices, complex micro/nanofabrication processes preclude the progress of extensive applications. We address this challenge by introducing porous polyethylene terephthalate (PET) membrane to develop a new type of nanotemplate electrode. This nanotemplate electrode is manufactured following a fabrication process on a porous PET membrane by atomic layer deposition. The 3D nanotemplate electrodes afford intracellular access to cardiomyocytes to report intracellular-like action potentials. These controllable nanotemplate electrodes exhibit sensitive and prolonged intracellular recordings of action potentials compared with free-growing 3D nanoelectrodes. This study indicates that the optimized structure of the nanoelectrode significantly promotes the performance of intracellular recording to assess electrophysiology in the fields of cardiology and neuroscience at an action potential level.

Intracellular Recording of Cardiomyocytes by Integrated Electrical Signal Recording and Electrical Pulse Regulating System.

The electrophysiological signal can reflect the basic activity of cardiomyocytes, which is often used to study the working mechanism of heart. Intracellular recording is a powerful technique for studying transmembrane potential, proving a favorable strategy for electrophysiological research. To obtain high-quality and high-throughput intracellular electrical signals, an integrated electrical signal recording and electrical pulse regulating system based on nanopatterned microelectrode array (NPMEA) is developed in this work. Due to the large impedance of the electrode, a high-input impedance preamplifier is required. The high-frequency noise of the circuit and the baseline drift of the sensor are suppressed by a band-pass filter. After amplifying the signal, the data acquisition card (DAQ) is used to collect the signal. Meanwhile, the DAQ is utilized to generate pulses, achieving the electroporation of cells by NPMEA. Each channel uses a voltage follower to improve the pulse driving ability and isolates each electrode. The corresponding recording control software based on LabVIEW is developed to control the DAQ to collect, display and record electrical signals, and generate pulses. This integrated system can achieve high-throughput detection of intracellular electrical signals and provide a reliable recording tool for cell electro-physiological investigation.