Integrated functional neuronal network analysis of 3D silk-collagen scaffold-based mouse cortical culture

Bioengineered 3D tunable neuronal constructs are a versatile platform for study-ing neuronal network functions, offering numerous advantages over existing technologies and providing for the discovery of new biological insights. Func-tional neural networks can be evaluated using calcium imaging and quantitatively described using network science. This protocol includes instructions for fabri-cating protein-based composite scaffolds, 3D in vitro culture of embryonic mouse cortical neurons, virally induced expression of GCaMP6f, wide-field cal-cium imaging, and computational analysis with open-source software and custom MATLAB code. For complete details on the use and execution of this protocol, please refer to Dingle et al. (2020).

Automated summary

Bioengineered 3D tunable neuronal constructs offer advantages for studying neuronal network functions and can be evaluated and described using calcium imaging and network science. The protocol includes instructions for scaffold fabrication, cell culture, viral expression, calcium imaging, and computational analysis.