A flattened retina-eyecup preparation suitable for electrophysiological studies of neurons visualized with trans-scleral infrared illumination

We present an in vitro flattened retinal-scleral preparation suitable for electrophysiological studies from visually targeted amacrine and ganglion cells of the rabbit retina. In a newly designed superfusion chamber, the retinal-scleral tissue is stained with Azure B allowing for imaging of neurons in the ganglion cell layer with an infrared (IR)-sensitive CCD camera via trans-scleral IR illumination. Neurons can be visually identified and targeted for both extracellular and intracellular recordings made singly or in simultaneous pairs. The quality and stability of the recordings are excellent and the tissue remains viable for up to 10 h. This relatively simple preparation avoids the extensive surgical manipulations inherent to those based on isolated retinas or retinal slices. Moreover, the use of trans-scleral IR illumination rather than fluorescent dyes to visualize and target neurons allows for electrophysiological studies of the retina under controlled adaptational states including dark-adapted conditions.