Electrophysiological Analysis of Retinal Organoid Development Using 3D Microelectrodes of Liquid Metals

Abstract Despite of the substantial potential of human‐derived retinal organoids, the degeneration of retinal ganglion cells (RGCs) during maturation limits their utility in assessing the functionality of later‐born retinal cell subtypes. Additionally, conventional analyses primarily rely on fluorescent emissions, which limits the detection of actual cell functionality while risking damage to the 3D cytoarchitecture of organoids. Here, an electrophysiological analysis is presented to monitor RGC development in early to mid‐stage retinal organoids, and compare distinct features with fully‐mature mouse retina. This approach utilizes high‐resolution 3D printing of liquid‐metal microelectrodes, enabling precise targeting of specific inner retinal layers within organoids. The adaptable distribution and softness of these microelectrodes facilitate the spatiotemporal recording of inner retinal signals. This study not only demonstrates the functional properties of RGCs in retinal organoid development but also provides insights into their synaptic connectivity, reminiscent of fetal native retinas. Further comparison with fully‐mature mouse retina in vivo verifies the organoid features, highlighting the potential of early‐stage retinal organoids in biomedical research.