Electroporation of Whole-Mount Postnatal Rodent Retinas for Advanced Functional Assays.

To study gene function in regulating rodent retinal waves during development, an efficient method for gene delivery into whole-mount retinas is required while preserving circuit functionality for physiological studies. We present an optimized electroporation protocol for developing rodent retinal explants. The procedure includes the fabrication of horizontally aligned platinum electrodes and the placement of retinal explants between them to generate a uniform electric field for high transfection efficiency. The entire process-dissection and electroporation-can be completed within 1-2 h. Successful transfection is verified by fluorescence microscopy, and physiological assays such as patch-clamp recordings and live imaging can be performed within 1-4 days following electroporation. This rapid and reliable protocol enables functional analysis for a specific gene in regulating retinal waves and can be adapted to other organotypic slice cultures. Key features • Incorporates horizontally aligned platinum electrodes and enables cell type-specific promoters to drive gene expression for physiological studies. • Preserves retinal wave activity while markedly improving transfection efficiency in whole-mount postnatal rodent retinas. • Requires only 1-2 h from retinal dissection to electroporation. • Allows completion of functional experiments within four days after electroporation.