Developing a new tool capable of high-precision electrophysiological recording of a large network of electrogenic cells has long been an outstanding challenge in neurobiology and cardiology. Here, we combine nanoscale intracellular electrodes with complementary metal-oxide-semiconductor (CMOS) integrated circuits to realize a high-fidelity all-electrical electrophysiological imager for parallel intracellular recording at the network level. Our CMOS nanoelectrode array has 1,024 recording/stimulation 'pixels' equipped with vertical nanoelectrodes, and can simultaneously record intracellular membrane potentials from hundreds of connected in vitro neonatal rat ventricular cardiomyocytes. We demonstrate that this network-level intracellular recording capability can be used to examine the effect of pharmaceuticals on the delicate dynamics of a cardiomyocyte network, thus opening up new opportunities in tissue-based pharmacological screening for cardiac and neuronal diseases as well as fundamental studies of electrogenic cells and their networks.

Current electrophysiological or optical techniques cannot reliably perform simultaneous intracellular recordings from more than a few tens of neurons. Here, we report a nanoelectrode array that can simultaneously obtain intracellular recordings from thousands of connected mammalian neurons in vitro. The array consists of 4,096 platinum-black electrodes with nanoscale roughness fabricated on top of a silicon chip that monolithically integrates 4,096 microscale amplifiers, configurable into pseudo-current-clamp mode (for concurrent current injection and voltage recording) or into pseudo-voltage-clamp mode (for concurrent voltage application and current recording). We used the array in pseudo-voltage-clamp mode to measure the effects of drugs on ion-channel currents. In pseudo-current-clamp mode, the array recorded intracellular action potentials and post-synaptic potentials from over thousands of neurons. In addition, we mapped over 300 excitatory and inhibitory synaptic connections from over 1,700 neurons that were recorded for 19 mins. This high-throughput intracellular-recording technology could benefit functional connectome mapping, electrophysiological screening, and other functional interrogations of neuronal networks. The patch clamp electrode is celebrated for its high-sensitivity intracellular recording that can measure not only action potential (AP) propagation in neurons but also subthreshold events such as postsynaptic potentials (PSPs). Dense, parallel execution of such high-Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

Electrode technology for electrophysiology has a long history of innovation, with some decisive steps including the development of the voltage-clamp measurement technique by Hodgkin and Huxley in the 1940s and the invention of the patch clamp electrode by Neher and Sakmann in the 1970s. The high-precision intracellular recording enabled by the patch clamp electrode has since been a gold standard in studying the fundamental cellular processes underlying the electrical activities of neurons and other excitable cells. One logical next step would then be to parallelize these intracellular electrodes, since simultaneous intracellular recording from a large number of cells will benefit the study of complex neuronal networks and will increase the throughput of electrophysiological screening from basic neurobiology laboratories to the pharmaceutical industry. Patch clamp electrodes, however, are not built for parallelization; as for now, only ∼10 patch measurements in parallel are possible. It has long been envisioned that nanoscale electrodes may help meet this challenge. First, nanoscale electrodes were shown to enable intracellular access. Second, because their size scale is within the normal reach of the standard top-down fabrication, the nanoelectrodes can be scaled into a large array for parallelization. Third, such a nanoelectrode array can be monolithically integrated with complementary metal-oxide semiconductor (CMOS) electronics to facilitate the large array operation and the recording of the signals from a massive number of cells. These are some of the central ideas that have motivated the research activity into nanoelectrode electrophysiology, and these past years have seen fruitful developments. This Account aims to synthesize these findings so as to provide a useful reference. Summing up from the recent studies, we will first elucidate the morphology and associated electrical properties of the interface between a nanoelectrode and a cellular membrane, clarifying how the nanoelectrode attains intracellular access. This understanding will be translated into a circuit model for the nanobio interface, which we will then use to lay out the strategies for improving the interface. The intracellular interface of the nanoelectrode is currently inferior to that of the patch clamp electrode; reaching this benchmark will be an exciting challenge that involves optimization of electrode geometries, materials, chemical modifications, electroporation protocols, and recording/stimulation electronics, as we describe in the Account. Another important theme of this Account, beyond the optimization of the individual nanoelectrode-cell interface, is the scalability of the nanoscale electrodes. We will discuss this theme using a recent development from our groups as an example, where an array of ca. 1000 nanoelectrode pixels fabricated on a CMOS integrated circuit chip performs parallel intracellular recording from a few hundreds of cardiomyocytes, which marks a new milestone in electrophysiology.

A CMOS-MEA device combined with new impedance and electrochemical techniques measures cell attachment, growth/wound healing, cell–cell adhesion, metabolic state, and redox properties with single-cell spatial resolution for cell-biology applications.

Precise quantification of plant nitrogen (N) nutrition status is essential for crop N management. The concept of critical N concentration (Nc) has been widely used for assessment of plant N status. This study aimed to develop a new winter wheat Nc dilution curve based on leaf area duration (LAD). Four field experiments were performed on different cultivars with different N fertilization modes in the Yangtze River basin and Yellow River basin in China. Results showed that the increase in LAD with increasing cumulative thermal time took the shape of an “S” type curve; whereas shoot N concentration decreased with increasing LAD, according to a power function. Both LAD and shoot N concentration increased with increasing N application. The new LAD based Nc dilution curve was determined and described as Nc = 1.6774 LAD−0.37 when LAD > 0.13. However, when LAD ≤ 0.13, Nc was constant and can be calculated by the equation when LAD = 0.13. The validation of Nc dilution curve with dataset acquired from independent experiments confirmed that N nutrition index (NNI) predictions based on the newly established Nc dilution curve could precisely diagnose N deficiency at different plant growth stages. The integrated N nutrition index (NNIinte), which was obtained by the weighted mean of NNI, was used to estimate shoot N concentration, shoot dry matter, LAD, and yield using regression functions. The linear relationships between NNIinte and these growth variables were well correlated. These results provided enough evidence that the new LAD–based Nc dilution curve could effectively and precisely diagnoses N deficiency in winter wheat crops.