Abstract
Photoconductive stimulation allows the noninvasive depolarization of neurons cultured on a silicon wafer. This technique relies on a beam of light to target a cell of interest while applying a voltage bias across the silicon wafer. The targeted cell is excited with minimal physiological manipulation, and, therefore, long-term modulation of activity patterns and investigations of biochemical mechanisms sensitive to physiological perturbations are possible. Ideologically similar to transistor-based neuronal interfaces, the photoconductive-stimulation method has the advantage of being able to extracellularly excite any neuron in a network regardless of its spatial position on the silicon substrate. This protocol can be easily implemented on a conventional reflected-light fluorescence microscope using materials and resources that are readily available. Time requirements are comparable to standard cell-culture and electrophysiology techniques. When combined with fluorescence imaging of various molecular probes, activity-dependent cellular processes can be dynamically monitored.
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Acknowledgements
We thank M. Sailor and B. Collins for their input on the design and troubleshooting of the stimulation device. This work was supported by grants from the National Institute on Drug Abuse (NIDA) and the National Institute of Mental Health (NIMH).
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Supplementary information
Supplementary Movie 1
Fluo-4 calcium imaging of neurons showing spontaneous activity and stimulus-triggered fluorescence changes. Stimulation at 2.5 Hz begins ∼19 s into the movie. The culture is the same as that illustrated in Figure 2. (AVI 1058 kb)
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Goda, Y., Colicos, M. Photoconductive stimulation of neurons cultured on silicon wafers. Nat Protoc 1, 461–467 (2006). https://doi.org/10.1038/nprot.2006.67
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DOI: https://doi.org/10.1038/nprot.2006.67
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