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Retinal prostheses have the potential to restore vision to blind patients that have retinitis pigmentosa or similar hereditary degenerative disorders, by electrically stimulating surviving retinal neurons through implanted electrode arrays. Current retinal prostheses provide limited visual acuity and one challenge is to spatially control neural activation following electrical stimulation. Most of the retinal prostheses are either epi-retinal - in front of the retinal ganglion cell layer, or sub-retinal - behind photoreceptor layer. In this study, we performed calcium imaging of ganglion cells from whole mounted retinas and compared the spread of neural activation between epi-retinal stimulation with a fiber electrode and sub-retinal stimulation with a disk electrode. We investigated the effects of phase durations on the spatial resolution of biphasic stimulation. Our results suggest that with fiber electrode epi-retinal stimulation, the axon bundles activation can lead to significant spread of stimulation, and cannot be avoided simply by changing the phase durations. However, sub-retinal stimulation with very short pulses (phase duration 0.033ms) can effectively confine the activation of retinal ganglion cells. more...