Fabrication and Characterization of Silicon-Based Antimonene Thin Film via Electron Beam Evaporation.

Antimonene has attracted much attention due to its excellent characteristics of high carrier mobility, thermoelectric properties and high stability. It has great application prospects in Q-switched lasers, laser protection and spintronics. At present, the epitaxy growth of antimonene mainly depends on molecular beam epitaxy. We have successfully prepared antimonene films on silicon, germanium/silicon substrates for the first time using electron beam evaporation coating and studied the effects of the deposition rate and substrate on the preparation of antimonene; film characterization was performed via confocal microprobe Raman spectroscopy, via X-ray diffraction and using a scanning electron microscope. Raman spectroscopy showed that different deposition rates can lead to the formation of different structures of antimonene, such as α phase and ß phase. At the same time, it was found that the growth of antimonene is also affected by different substrates and ion beams.

Elevating intracellular action potential recording in cardiomyocytes: A precision-enhanced and biosafe single-pulse electroporation system.

Action potentials play a pivotal role in diverse cardiovascular physiological mechanisms. A comprehensive understanding of these intricate mechanisms necessitates a high-fidelity intracellular electrophysiological investigative approach. The amalgamation of micro-/nano-electrode arrays and electroporation confers substantial advantages in terms of high-resolution intracellular recording capabilities. Nonetheless, electroporation systems typically lack precise control, and commonly employed electroporation modes, involving tailored sequences, may escalate cellular damage and perturbation of normal physiological functions due to the multiple or higher-intensity electrical pulses. In this study, we developed an innovative electrophysiological biosensing system customized to facilitate precise single-pulse electroporation. This advancement serves to achieve optimal and uninterrupted intracellular action potential recording within cardiomyocytes. The refinement of the single-pulse electroporation technique is realized through the integration of the electroporation and assessment biosensing system, thereby ensuring a consistent and reliable means of achieving stable intracellular access. Our investigation has unveiled that the optimized single-pulse electroporation technique not only maintains robust biosafety standards but also enables the continuous capture of intracellular electrophysiological signals across an expansive three-day period. The universality of this biosensing system, adaptable to various micro/nano devices, furnishes real-time analysis and feedback concerning electroporation efficacy, guaranteeing the sustained, secure, and high-fidelity acquisition of intracellular data, thereby propelling the field of cardiovascular electrophysiological research.

Dynamic and Label-Free Sensing of Cardiomyocyte Responses to Nanosized Vesicles for Cardiac Oxidative Stress Injury Therapy.

Cardiac oxidative stress is a significant phenotype of myocardial infarction disease, a leading cause of global health threat. There is an urgent need to develop innovative therapies. Nanosized extracellular vesicle (nEV)-based therapy shows promise, yet real-time monitoring of cardiomyocyte responses to nEVs remains a challenge. In this study, a dynamic and label-free cardiomyocyte biosensing system using microelectrode arrays (MEAs) was constructed. Cardiomyocytes were cultured on MEA devices for electrophysiological signal detection and treated with nEVs from E. coli, gardenia, HEK293 cells, and mesenchymal stem cells (MSC), respectively. E. coli-nEVs and gardenia-nEVs induced severe paroxysmal fibrillation, revealing distinct biochemical communication compared to MSC-nEVs. Principal component analysis identified variations and correlations between nEV types. MSC-nEVs enhanced recovery without inducing arrhythmias in a H2O2-induced oxidative stress injury model. This study establishes a fundamental platform for assessing biochemical communication between nEVs and cardiomyocytes, offering new avenues for understanding nEVs' functions in the cardiovascular system.

Optimizing the Cell-Nanostructure Interface: Nanoconcave/Nanoconvex Device for Intracellular Recording of Cardiomyocytes.

Nanostructures are powerful components for the development of high-performance nanodevices. Revealing and understanding the cell-nanostructure interface are essential for improving and guiding nanodevice design for investigations of cell physiology. For intracellular electrophysiological detection, the cell-nanostructure interface significantly affects the quality of recorded intracellular action potentials and the application of nanodevices in cardiology research and pharmacological screening. Most of the current investigations of biointerfaces focus on nanovertical structures, and few involve nanoconcave structures. Here, we design both nanoconvex and nanoconcave devices to perform intracellular electrophysiological recordings. The amplitude, signal-to-noise ratio, duration, and repeatability of the recorded intracellular electrophysiological signals provide a multifaceted characterization of the cell-nanostructure interface. We demonstrate that devices based on both convex and concave nanostructures can create tight coupling, which facilitates high-quality and stable intracellular recordings and paves the way for precise electrophysiological study.

Three-Dimensional Cardiomyocyte-Nanobiosensing System for Specific Recognition of Drug Subgroups.

Abnormal cardiac electrophysiological activities significantly contribute to the incidence of cardiovascular diseases. Therefore, it is crucial to recognize effective drugs, which require an accurate, stable, and sensitive platform. Although conventional extracellular recordings offer a non-invasive and label-free manner to monitor the electrophysiological state of cardiomyocytes, the misrepresented and low-quality extracellular action potentials are difficult to provide accurate and high-content information for drug screening. This study presents the development of a three-dimensional cardiomyocyte-nanobiosensing system that can specifically recognize drug subgroups. The nanopillar-based electrode is manufactured by template synthesis and standard microfabrication technology on a porous polyethylene terephthalate membrane. Based on the cardiomyocyte-nanopillar interface, high-quality intracellular action potentials can be recorded by the minimally invasive electroporation. We validate the performance of a cardiomyocyte-nanopillar-based intracellular electrophysiological biosensing platform by two subclasses of sodium channel blockers, quinidine and lidocaine. The recorded intracellular action potentials accurately reveal the subtle differences between these drugs. Our study indicates that high-content intracellular recordings utilizing nanopillar-based biosensing can provide a promising platform for the electrophysiological and pharmacological investigation of cardiovascular diseases.

Integrated Cardiomyocyte-Based Biosensing Platform for Electroporation-Triggered Intracellular Recording in Parallel with Delivery Efficiency Evaluation.

Electroporation is a proven technique that can record action potential of cardiomyocytes and serve for biomolecular delivery. To ensure high cell viability, micro-nanodevices cooperating with low-voltage electroporation are frequently utilized in research, and the effectiveness of delivery for intracellular access is typically assessed using an optical imaging approach like flow cytometry. However, the efficiency of in situ biomedical studies is hampered by the intricacy of these analytical approaches. Here, we develop an integrated cardiomyocyte-based biosensing platform to effectively record action potential and evaluate the electroporation quality in terms of viability, delivery efficiency, and mortality. The ITO-MEA device of the platform possesses sensing/stimulating electrodes which combines with the self-developed system to achieve intracellular action potential recording and delivery by electroporation trigger. Moreover, the image acquisition processing system analyzes various parameters effectively to assess delivery performance. Therefore, this platform has the potential for drug delivery therapy and pathology research for cardiology.

Scalable and Robust Hollow Nanopillar Electrode for Enhanced Intracellular Action Potential Recording.

Electrophysiology is a unique biomarker of the electrogenic cells that can perform a disease investigation or drug assessment. In the recent decade, vertical nanoelectrode arrays can successfully achieve a high-quality intracellular electrophysiological study in electrogenic cells and their networks. However, a high success rate and high-quality and long-term intracellular recording using low-cost nanostructures is still a considerable challenge. Herein, we develop a scalable and robust hollow nanopillar electrode to achieve enhanced intracellular recording of cardiomyocytes. The template-based synthesis of vertical hollow nanopillars is compatible with large-scale and efficient microfabrication processes and is convenient to regulate the geometry of hollow nanopillars. Compared with the conventional same-size planar electrode, the regulating height of a hollow nanopillar can achieve high-quality and prolonged intracellular recordings, which can improve the cell-electrode interface for tight coupling and effective electroporation. It is demonstrated that the geometry regulation of a nanostructure is a powerful strategy to enhance intracellular recording.

Research Progress of Wide Tunable Bragg Grating External Cavity Semiconductor Lasers.

In this paper, we review the progress of wide tunable Bragg grating external cavity semiconductor lasers (BG-ECSLs). We concentrate on BG-ECSLs based on the wide tunable range for multicomponent detection. Wide tunable BG-ECSLs have many important applications, such as wavelength-division multiplexing (WDM) systems, coherent optical communications, gas detection and atom cooling. Wide tunability, narrow linewidth and a high side-mode suppression ratio BG-ECSLs have attracted much attention for their merits. In this paper, three main structures for achieving widely tunable, narrow linewidth, high side-mode suppression ratio BG-ECSLs are reviewed and compared in detail, such as the volume Bragg grating (VBG) structure, fiber Bragg grating (FBG) structure and waveguide Bragg grating (WBG) structure of ECSLs. The advantages and disadvantages of different structures of BG-ECSLs are analyzed. The results show that WBG-ECSLs are a potential way to realize the integration, small size, wide tuning range, stable spectral output and high side-mode suppression ratio laser output. Therefore, the use of WBG as optical feedback elements is still the mainstream direction of BG-ECSLs, and BG-ECSLs offer a further new option for multicomponent detection and multi-atoms cooling.

Scalable Nanotrap Matrix Enhanced Electroporation for Intracellular Recording of Action Potential.

Electrophysiological recording, as a long-sought objective, plays a crucial role in fundamental biomedical research and practical clinical applications. The challenge in developing electrophysiological detection platforms is to combine simplicity, stability, and sensitivity in the same device. In this study, we develop a nanotrapped microelectrode based on a porous PET membrane, which is compatible with large-scale microtechnologies. The nanotraps can promote the protrusion of the local cell membrane in the hollow center and offer a unique nanoedge structure for tight sealing and effective electroporation. We demonstrate that scalable nanotraps can enhance cell-electrode coupling and perform high-quality intracellular recording. Further, the nanoedge-enhanced electroporation and minimally invasive nanotrapped recordings afford much longer intracellular access of over 100 min and permit consecutive electroporation events in a short period of time. This study suggests that the geometry-regulating strategy of the cell-electrode nanointerface could significantly improve the intracellular recording performance of a nanopatterned electrode.

Universal and Sensitive Drug Assessment Biosensing Platform Using Optimal Mechanical Beating Detection of Single Cardiomyocyte.

The preclinical assessment of efficacy and safety is essential for cardiovascular drug development in order to guarantee effective prevention and treatment of cardiovascular disease and avoid human health endangerment and a huge waste of resources. Rhythmic mechanical beating as one of the crucial cardiomyocyte properties has been exploited to establish a drug assessment biosensing platform. However, the conventional label-free biosensing platforms are difficult to perform high-throughput and high-resolution mechanical beating detection for a single cardiomyocyte, while label-based strategies are limited by pharmacologically adverse effects and phototoxicity. Herein, we propose a biosensing platform involving the multichannel electrode array device and the universal mechanical beating detection system. The platform can determine the optimal characteristic working frequency of different devices and dynamically interrogate the viability of multisite single cardiomyocytes to establish the optimized cell-based model for sensitive drug assessment. The subtle changes of mechanical beating signals induced by cardiovascular drugs can be detected by the platform, thereby demonstrating its high performance in pharmacological assessment. The universal and sensitive drug assessment biosensing platform is believed to be widely applied in cardiology investigating and preclinical drug screening.

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.

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.

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.

Integrated Au-Nanoroded Biosensing and Regulating Platform for Photothermal Therapy of Bradyarrhythmia.

Bradyarrhythmia is a kind of cardiovascular disease caused by dysregulation of cardiomyocytes, which seriously threatens human life. Currently, treatment strategies of bradyarrhythmia mainly include drug therapy, surgery, or implantable cardioverter defibrillators, but these strategies are limited by drug side effect, surgical trauma, and instability of implanted devices. Here, we developed an integrated Au-nanoroded biosensing and regulating platform to investigate the photothermal therapy of cardiac bradyarrhythmia in vitro. Au-nanoroded electrode array can simultaneously accumulate energy from the photothermal regulation and monitor the electrophsiological state to restore normal rhythm of cardiomyocytes in real time. To treat the cardiomyocytes cultured on Au-nanoroded device by near-infrared (NIR) laser irradiation, cardiomyocytes return to normal for long term after irradiation of suitable NIR energy and maintenance. Compared with the conventional strategies, the photothermal strategy is more effective and convenient to regulate the cardiomyocytes. Furthermore, mRNA sequencing shows that the differential expression genes in cardiomyocytes are significantly increased after photothermal strategy, which are involved in the regulation of the heart rate, cardiac conduction, and ion transport. This work establishes a promising integrated biosensing and regulating platform for photothermal therapy of bradyarrhythmia in vitro and provides reliable evidence of photothermal regulation on cardiomyocytes for cardiological clinical studies.

A dynamic and quantitative biosensing assessment for electroporated membrane evolution of cardiomyocytes.

The electrophysiological study is an essential approach to perform the biology and basic medicine research. To achieve the intracellular electrophysiological investigation, electroporation is introduced as an effective and convenient strategy to achieve the intracellular access of electrogenic cells and obtain high-fidelity action potentials. However, seldom platform could provide a quantitative and dynamic strategy to assess the electroporation-induced membrane perforation and recovery during intracellular electrophysiological investigation. Here we develop a high-throughput, sensitive, and stable biosensing platform to assess the evolution of electroporated cell membrane dynamically and quantitatively based on the recorded intracellular electrophysiological signals of cardiomyocytes. Following the electroporation, the extracellular action potentials transiently convert to the intracellular action potentials, whose amplitude rapidly increases to the maximum and then gradually decays. The intracellular action potentials finally convert back to the extracellular action potentials. This biosensing platform can dynamically explore and characterize the evolution procedures of perforation, stabilization, and resealing of the cell membrane by intracellular recordings. Moreover, the effect of electroporation voltages on the cell membrane is segmentally and quantitatively analyzed, demonstrating that a higher electroporation voltage induced a longer resealing time within the safe range of electroporation voltage. We believed that this dynamic and quantitative electroporated membrane evolution biosensing assessment platform will be a promising tool to pave a new avenue to bridge the electrophysiology and electroporated membrane evolution.

Flexible Tongue Electrode Array System for In Vivo Mapping of Electrical Signals of Taste Sensation.

Tongue is a unique organ that senses tastes, and the scientific puzzle about whether electricity can evoke taste sensations and how the sensations have been distributed on the tongue has not been solved. Investigations on tongue stimulation by electricity might benefit the developments of techniques for clinical neuromodulation, tissue activation, and a brain-tongue-machine interface. To solve the scientific puzzle of whether electrical stimulation induces taste-related sensations, a portable flexible tongue electrode array system (FTEAS) was developed, which can synchronously provide electrical stimulation and signal mapping at each zone of the tongue. Utilizing the FTEAS to perform tests on the rat tongue in vivo, specific electrical signals were observed to be evoked by chemical and electrical stimulations. The features and distributions of the electric signals evoked during the rat tongue tests were systematically studied and comprehensively analyzed. The results show that an appropriate electrical stimulation can induce multiple sensations simultaneously, while the distribution of each sensation was not significantly distinguished among different zones of the tongue, and at the same time, this taste-related electrical signal can be recorded by the FTEAS. This work establishes a promising platform to solve the scientific puzzle of how sensations are activated chemically and electrically on the tongue and may provide advanced noninvasive oral-electrotherapy and a brain-tongue-machine interface.

Cardiomyocyte electrical-mechanical synchronized model for high-content, dose-quantitative and time-dependent drug assessment.

Cardiovascular diseases have emerged as a significant threat to human health. However, drug development is a time-consuming and costly process, and few drugs pass the preclinical assessment of safety and efficacy. The existing patch-clamp, Ca2+ imaging, and microelectrode array technologies in cardiomyocyte models for drug preclinical screening have suffered from issues of low throughput, limited long-term assessment, or inability to synchronously and correlatively analyze electrical and mechanical signals. Here, we develop a high-content, dose-quantitative and time-dependent drug assessment platform based on an electrical-mechanical synchronized (EMS) biosensing system. This microfabricated EMS can record both firing potential (FP) and mechanical beating (MB) signals from cardiomyocytes and extract a variety of characteristic parameters from these two signals (FP-MB) for further analysis. This system was applied to test typical ion channel drugs (lidocaine and isradipine), and the dynamic responses of cardiomyocytes to the tested drugs were recorded and analyzed. The high-throughput characteristics of the system can facilitate simultaneous experiments on a large number of samples. Furthermore, a database of various cardiac drugs can be established by heat map analysis for rapid and effective screening of drugs. The EMS biosensing system is highly promising as a powerful tool for the preclinical development of new medicines.

Accurate and efficient intracellular delivery biosensing system by nanostrawed electroporation array.

Electroporation serves as a powerful technique to introduce the exogenous nucleotides, DNA, RNA, proteins, dyes, and virus particles into cells. Through the effect of high intensity of electric field, the permeability of the cell membrane is instantaneously improved to absorb the exogenous molecules in surrounding medium. To protect the cell viability, ultralow-voltage electroporation techniques are well developed by versatile devices, and delivery efficiency is commonly assessed by label-based analysis by microscope-ImageJ or flow cytometry. However, accuracy and complexity of these analytical strategies still hinder efficient and precise biomedical studies in situ. Here we developed an intracellular delivery biosensing system by nanostrawed electroporation array (NEA) that can efficiently assess the universal electroporation performance by cell viability, delivery efficiency, and cell mortality. The intracellular delivery biosensing system consists of NEA, fluorescent microscope, and automated analysis software. Intracellular delivery biosensing system of electroporation quality is based on the enhanced fluorescent watershed segmentation (enhanced FWS) algorithm, which possessed low deviation (~5%) and significantly shortened the operation time (~8 s/10 images) in contrast to high deviation (~13%) and long operation time (~10 min/10 images) of conventional inaccurate, labor-intensive and time-consuming methods. It is believed that the intracellular delivery biosensing system will be a promising universal platform to assess nanodevice electroporation in the fields of cell biology, biotechnology, and medicine.

Synchronized intracellular and extracellular recording of action potentials by three-dimensional nanoroded electroporation.

Electrophysiological study is an essential and significant strategy to explore the biological mechanism of electrogenic cells. Current advanced nanodevices can achieve the high-fidelity intracellular electrophysiological recordings, and most of detection systems record the extracellular and intracellular action potentials (EAPs and IAPs) in an asynchronous or isolated manner, so it is demanded to develop the platform to reveal correlation between EAP and IAP recording. Here, we establish a utility strategy to achieve synchronized intracellular and extracellular recording of neonatal rat cardiomyocytes by low-voltage three-dimensional (3D) nanoroded electroporation. By integrating the advantages of nanodevice and microdevice, 3D nanoroded microdevice is developed to achieve the high-throughput large-scale synchronous intracellular and extracellular electrophysiological study. By applying low-voltage electroporation, intracellular and extracellular signals can be synchronously acquired from intracellular access and extracellular coupling, respectively. Recorded synchronized signals contain both typical EAPs and IAPs, which have good synchronicity in spatiotemporal dimensions at each recording site. Moreover, correlation between both signals is further bridged in experimental and simulated way. This intracellular electrophysiological platform presents unique advantages over the conventional system to achieve the synchronized intracellular and extracellular electrophysiological study at membrane voltage level.

In-Cell Nanoelectronics: Opening the Door to Intracellular Electrophysiology.

Establishing a reliable electrophysiological recording platform is crucial for cardiology and neuroscience research. Noninvasive and label-free planar multitransistors and multielectrode arrays are conducive to perform the large-scale cellular electrical activity recordings, but the signal attenuation limits these extracellular devices to record subthreshold activities. In recent decade, in-cell nanoelectronics have been rapidly developed to open the door to intracellular electrophysiology. With the unique three-dimensional nanotopography and advanced penetration strategies, high-throughput and high-fidelity action potential like signal recordings is expected to be realized. This review summarizes in-cell nanoelectronics from versatile nano-biointerfaces, penetration strategies, active/passive nanodevices, systematically analyses the applications in electrogenic cells and especially evaluates the influence of nanodevices on the high-quality intracellular electrophysiological signals. Further, the opportunities, challenges and broad prospects of in-cell nanoelectronics are prospected, expecting to promote the development of in-cell electrophysiological platforms to meet the demand of theoretical investigation and clinical application.