Improving reliability and reducing costs of cardiotoxicity assessments using laser-induced cell poration on microelectrode arrays.

Drug-induced cardiotoxicity is a major barrier to drug development and a main cause of withdrawal of marketed drugs. Drugs can strongly alter the spontaneous functioning of the heart by interacting with the cardiac membrane ion channels. If these effects only surface during in vivo preclinical tests, clinical trials or worse after commercialization, the societal and economic burden will be significant and seriously hinder the efficient drug development process. Hence, cardiac safety pharmacology requires in vitro electrophysiological screening assays of all drug candidates to predict cardiotoxic effects before clinical trials. In the past 10 years, microelectrode array (MEA) technology began to be considered a valuable approach in pharmaceutical applications. However, an effective tool for high-throughput intracellular measurements, compatible with pharmaceutical standards, is not yet available. Here, we propose laser-induced optoacoustic poration combined with CMOS-MEA technology as a reliable and effective platform to detect cardiotoxicity. This approach enables the acquisition of high-quality action potential recordings from large numbers of cardiomyocytes within the same culture well, providing reliable data using single-well MEA devices and single cardiac syncytia per each drug. Thus, this technology could be applied in drug safety screening platforms reducing times and costs of cardiotoxicity assessments, while simultaneously improving the data reliability.

Use of a CD laser pickup head to fabricate microelectrodes in polymethylmethacrylate substrates for biosensing applications.

In this work, we report a simple fabrication method for microelectrodes on a polymethylmethacrylate substrate, using a low-cost laser platform based on a CD-DVD unit for direct rapid-prototyping. We used this laser microfabrication technique to etch any desired design on polymethylmethacrylate substrates to produce microchannels with controlled geometry, with a highly repeatable micron-scale resolution. Those shallow microchannels were then filled with a conductive paste of material of our choice that was converted into microelectrodes of desired shapes and geometries after drying. To validate our process, different geometries, sizes and materials were used as electrodes, and then tested for amperometry and impedance measurements. Development of these microelectrodes is motivated by their potential application in sensors and biosensors, such as glucose and cell counting, as demonstrated in this paper.

A low-cost, multiplexed µECoG system for high-density recordings in freely moving rodents.

OBJECTIVE: Micro-electrocorticography (µECoG) offers a minimally invasive neural interface with high spatial resolution over large areas of cortex. However, electrode arrays with many contacts that are individually wired to external recording systems are cumbersome and make recordings in freely behaving rodents challenging. We report a novel high-density 60-electrode system for µECoG recording in freely moving rats. APPROACH: Multiplexed headstages overcome the problem of wiring complexity by combining signals from many electrodes to a smaller number of connections. We have developed a low-cost, multiplexed recording system with 60 contacts at 406 µm spacing. We characterized the quality of the electrode signals using multiple metrics that tracked spatial variation, evoked-response detectability, and decoding value. Performance of the system was validated both in anesthetized animals and freely moving awake animals. MAIN RESULTS: We recorded µECoG signals over the primary auditory cortex, measuring responses to acoustic stimuli across all channels. Single-trial responses had high signal-to-noise ratios (SNR) (up to 25 dB under anesthesia), and were used to rapidly measure network topography within ∼10 s by constructing all single-channel receptive fields in parallel. We characterized evoked potential amplitudes and spatial correlations across the array in the anesthetized and awake animals. Recording quality in awake animals was stable for at least 30 days. Finally, we used these responses to accurately decode auditory stimuli on single trials. SIGNIFICANCE: This study introduces (1) a µECoG recording system based on practical hardware design and (2) a rigorous analytical method for characterizing the signal characteristics of µECoG electrode arrays. This methodology can be applied to evaluate the fidelity and lifetime of any µECoG electrode array. Our µECoG-based recording system is accessible and will be useful for studies of perception and decision-making in rodents, particularly over the entire time course of behavioral training and learning.

Use of screen-printed microelectrodes working as generator/collector systems for the determination of the antioxidant capacity of phenolic compounds.

A new method using cheap homemade dual-electrodes has been developed to measure the antioxidant capacity of phenolic compounds. These micro-sized electrodes are elaborated by successive screen-printing of conductive ink and insulator layers and are then used as generator/collector sensors. Cyclic voltammetry and chronoamperometry with a bipotentiostat have been used to test and characterize these sensors. The antioxidant capacity values found by this new method are compared with a classical method (using a macro-sized electrode) to demonstrate its reliability.

A cost analysis of intraoperative microelectrode recording during subthalamic stimulation for Parkinson's disease.

Deep brain stimulation of the subthalamic nucleus is the standard of care for treating medically intractable Parkinson's disease. Although the adjunct of microelectrode recording improves the targeting accuracy of subthalamic nucleus deep brain stimulation in comparison with image guidance alone, there has been no investigation of the financial cost of intraoperative microelectrode recording. This study was performed to address this issue. A comprehensive literature search of large subthalamic nucleus deep brain stimulation series (minimum, 75 patients) was performed, revealing a mean operating room time of 223.83 minutes for unilateral and 279.79 minutes for simultaneous bilateral implantation. The baseline operating room time was derived from the published operating room time for subthalamic nucleus deep brain stimulation without microelectrode recording. The total cost (operating room, anesthesia, neurosurgery) was then calculated based on hospitals geographically representative of the entire United States. The average cost for subthalamic nucleus deep brain stimulation implantation with microelectrode recording per patient is $26,764.79 for unilateral, $33,481.43 for simultaneous bilateral, and $53,529.58 for staged bilateral. For unilateral implantation, the cost of microelectrode recording is $19,461.75, increasing the total cost by 267%. For simultaneous bilateral implantation, microelectrode recording costs $20,535.98, increasing the total cost by 159%. For staged bilateral implantation, microelectrode recording costs $38,923.49, increasing the total cost by 267%. Microelectrode recording more than doubles the cost of subthalamic nucleus deep brain stimulation for Parkinson's disease and more than triples the cost for unilateral and staged bilateral procedures. The cost burden of microelectrode recording to subthalamic nucleus deep brain stimulation requires the clinical efficacy of microelectrode recording to be proven in a prospective evidence-based manner in order to curtail the potential for excessive financial burden to the health care system.

An inexpensive drivable cannulated microelectrode array for simultaneous unit recording and drug infusion in the same brain nucleus of behaving rats.

Neurons are functionally segregated into discrete populations that perform specific computations. These computations, mediated by neuron-neuron electrochemical signaling, form the neural basis of behavior. Thus fundamental to a brain-based understanding of behavior is the precise determination of the contribution made by specific neurotransmitters to behaviorally relevant neural activity. To facilitate this understanding, we have developed a cannulated microelectrode array for use in behaving rats that enables simultaneous neural ensemble recordings and local infusion of drugs in the same brain nucleus. The system is inexpensive, easy to use, and produces robust and quantitatively reproducible drug effects on recorded neurons.

Quantifying the benefits of additional channels of multifocal VEP recording.

For some individuals and for some locations, multifocal visual evoked potentials (mfVEP) may be too small or appear 'too noisy' to be reliably measured. By adding electrodes, especially electrodes placed lateral to the midline, and by recording with multiple channels, the amplitude of the signal can be increased in some field locations. However, the addition of electrodes involves certain costs; the set-up time is longer and the data analysis more time consuming and complex. The objective of this study was to assess the benefits of adding electrodes by quantifying these benefits using a signal-to-noise measure. In addition to the typical midline placement of electrodes, two electrodes were placed 1 cm above and 4 cm lateral to the inion on each side. This allowed for 3 channels of recording and 3 additional, derived channels. The mfVEPs were recorded with a 60 sector, pattern-reversing display presented to one eye. Two 7 min records were obtained from 14 individuals with no known visual problems. The two records were averaged and a signal-to-noise (SNR) measure was obtained for every response from all 6 channels. For each sector of the display and each subject, the benefits of additional electrodes were quantified by comparing the SNR from the traditional midline channel to the best SNR from amongst the 6 channels. The number of responses exceeding any given criterion SNR value was increased with the additional channels. For example, 79% of the responses for the typical midline channel exceeded a SNR of 0.6 (a false positive rate of about 2.5%) and this increased to 93% when the best SNR value was used. As expected, summing the mfVEP responses from contiguous sectors also increased the SNR values. Additional electrodes and multiple channels of recording substantially improve the quality of the mfVEP records and the SNR measure provides a useful metric for assessing these benefits.