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This Protocol Extension describes the low-cost production of rapidly customizable optical neural probes for in vivo optogenetics. We detail the use of a 3D printer to fabricate minimally invasive microscale inorganic LED-based neural probes which can control neural circuit activity in freely behaving animals, thus extending the scope of two previously published protocols describing the fabrication and implementation of optoelectronic devices for studying intact neural systems. The 3D-printing fabrication process does not require extensive training and eliminates the need for expensive materials, specialized cleanroom facilities, and time-consuming microfabrication techniques typical of conventional manufacturing processes. As a result, the design of the probes can be quickly optimized, based on experimental need, reducing the cost and turnaround for customization. For example, 3D-printed probes can be customized to target multiple brain regions or scaled up for use in large animal models. This protocol comprises three procedures: 1) probe fabrication, 2) wireless module preparation, and 3) implantation for in vivo assays. For experienced researchers, neural probe and wireless module fabrication requires approximately two days, while implantation should take 30–60 minutes per animal. Time required for behavioural assays will vary depending on the experimental design and should include at least 5 days of animal handling prior to the implantation of the probe, to familiarize each animal to their handler, thus reducing handling stress which may influence the result of the behavioural assays. The implementation of customized probes improves the flexibility in optogenetic experimental design and increases access to wireless probes for in vivo optogenetic research. |
