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Processes 2017, 5(2), 30; doi:10.3390/pr5020030

Closed-Loop Characterization of Neuronal Activation Using Electrical Stimulation and Optical Imaging

1
School of Electrical and Computer Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA
2
Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA
3
Laboratory for Neuroengineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA
4
School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Michael Henson
Received: 22 January 2017 / Revised: 30 April 2017 / Accepted: 26 May 2017 / Published: 3 June 2017
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Abstract

We have developed a closed-loop, high-throughput system that applies electrical stimulation and optical recording to facilitate the rapid characterization of extracellular, stimulus-evoked neuronal activity. In our system, a microelectrode array delivers current pulses to a dissociated neuronal culture treated with a calcium-sensitive fluorescent dye; automated real-time image processing of high-speed digital video identifies the neuronal response; and an optimized search routine alters the applied stimulus to achieve a targeted response. Action potentials are detected by measuring the post-stimulus, calcium-sensitive fluorescence at the neuronal somata. The system controller performs directed searches within the strength–duration (SD) stimulus-parameter space to build probabilistic neuronal activation curves. This closed-loop system reduces the number of stimuli needed to estimate the activation curves when compared to the more commonly used open-loop approach. This reduction allows the closed-loop system to probe the stimulus regions of interest in the multi-parameter waveform space with increased resolution. A sigmoid model was fit to the stimulus-evoked activation data in both current (strength) and pulse width (duration) parameter slices through the waveform space. The two-dimensional analysis results in a set of probability isoclines corresponding to each neuron–electrode pair. An SD threshold model was then fit to the isocline data. We demonstrate that a closed-loop methodology applied to our imaging and micro-stimulation system enables the study of neuronal excitation across a large parameter space. View Full-Text
Keywords: extracellular electrical stimulation; closed-loop; strength-duration; micro-electrode array (MEA); dissociated culture; activation curve; optical recording extracellular electrical stimulation; closed-loop; strength-duration; micro-electrode array (MEA); dissociated culture; activation curve; optical recording
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Kuykendal, M.L.; Guvanasen, G.S.; Potter, S.M.; Grover, M.A.; DeWeerth, S.P. Closed-Loop Characterization of Neuronal Activation Using Electrical Stimulation and Optical Imaging. Processes 2017, 5, 30.

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