This article describes the protocol for culturing low-density primary hippocampal neurons growing on glass coverslips inverted over a glial monolayer. The neuron and glial layers are separated by paraffin wax beads. The neurons grown by this method are suitable for high-resolution optical imaging and functional assays.
Roppongi, R. T., Champagne-Jorgensen, K. P., Siddiqui, T. J. Low-Density Primary Hippocampal Neuron Culture. J. Vis. Exp. (122), e55000, doi:10.3791/55000 (2017).