Optoretinogram, a technique in which optical coherence tomography (OCT) is used to measure retinal functions in response to a visible light stimulus, can be a potentially useful tool to quantify retinal health alterations. Existing experimental studies on animals have focused on measuring the global retinal response by transversally averaging 3D data across the retina, which minimizes the spatial resolution of the signals, and limits the signal-to-noise ratio (SNR) because only central B-scans are collected and analyzed. These problems were addressed in this study by collecting volumetric data to probe functional signals and developing an improved 3D registration approach to align such series-acquired OCT volumes. These data were then divided into small blocks and subject to a spatiotemporal analysis, whose results confirmed the spatial-dependence of functional signals. By further averaging, the overall measurement accuracy for position and scattering signals was estimated to be approximately 30 nm and 1.1%, respectively. With improved accuracy, this method revealed certain novel functional signals that have not been previously reported. In conclusion, this work provides a powerful tool to monitor retinal local and global functional changes in aging, diseased, or treated rodent eyes.

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