PT  - JOURNAL ARTICLE
AU  - Joseph Negri
AU  - Vilas Menon
AU  - Tracy L. Young-Pearse
TI  - Assessment of spontaneous neuronal activity <em>in vitro</em> using multi-well multi-electrode arrays: Implications for assay development
AID  - 10.1523/ENEURO.0080-19.2019
DP  - 2020 Jan 02
TA  - eneuro
PG  - ENEURO.0080-19.2019
4099  - http://www.eneuro.org/content/early/2020/01/02/ENEURO.0080-19.2019.short
4100  - http://www.eneuro.org/content/early/2020/01/02/ENEURO.0080-19.2019.full
AB  - Multi-electrode arrays (MEA) are being more widely used by researchers as an instrument platform for monitoring prolonged, non-destructive recordings of spontaneously firing neurons in vitro for applications in modeling Alzheimer’s, Parkinson’s, schizophrenia, and many other diseases of the human central nervous system. With the more widespread use of these instruments, there is a need to examine the prior art of studies utilizing MEAs, and delineate best practices for data acquisition and analysis to avoid errors in interpretation of the resultant data. Using a dataset of recordings from primary rat (Rattus norvegicus) cortical cultures, methods and statistical power for discerning changes in neuronal activity on the array level are examined. Further, a method for unsupervised spike sorting is implemented, allowing for the resolution of action potential incidents down to the single neuron level. Following implementation of spike sorting, the dynamics of firing frequency across populations of individual neurons and networks are examined longitudinally. Finally, the ability to detect a frequency independent phenotype, the change in action potential amplitude, is demonstrated through the use of pore-forming neurotoxin treatments. Taken together, this study provides guidance and tools for users wishing to incorporate multi-well MEA usage into their studies.Significance Statement Multi-electrode arrays (MEA) are an instrument platform being utilized by an increasing number of neuroscientist for the purpose of monitoring spontaneous firing of neurons in vitro over extended periods of time. Through an analysis of existing literature and empirically generated datasets, this study seeks to establish best practices for the use of these instruments for applications employing neuronal cultures. Elements of experimental design and analysis for assaying firing frequencies across arrays are discussed, with a focus on the use of multi-well MEAs. Additionally, methods for (i) resolving signal to individual neurons through unsupervised spike-sorting, (ii) assessing network dynamics, and (iii) quantifying changing in action potential amplitudes are reported.