The optical recording of light-evoked activity in populations of neurons in the mammalian retina offers several benefits over the use of multielectrode arrays. However, population imaging has been hindered by the effective loading of synthetic fluorescent indicators, especially in the mature tissue. We have therefore developed an electroporation method to label the complete ganglion cell layer of the adult mammalian retina. We optimized the protocol such that the retina recovers from electroporation and generates responses to visual stimuli. The method can be used with a diverse set of indicators with a range of affinities and emission wavelengths. It therefore can be combined with transgenic animals expressing fluorescent markers to target specific neuronal types. Importantly, the ganglion cell layer remains accessible for subsequent intracellular recording and morphological identification.