The study of cell morphology is important and prevalent in understanding normal and pathological conditions in the brain. Brain cells are common targets for treatment for brain diseases. Specifically, microglia – the brain’s resident immune cells – undergo a range of morphological changes in response to injury and are targets of many mitigating treatments. Image processing has been a valuable tool to assess microglial cell morphology via the analysis of microglial shape features and there continue to be additional opportunities for further investigation. Prior research has indicated a connection between features such as solidity and extent, two shape features that measure the ratio of cell areas. In this study, we examine three shape features of fluorescently labeled microglia: Euler number, extent, and solidity, in the context of ischemic injury. Ischemic injury was modeled using oxygen-glucose deprivation (OGD) in cultured whole hemisphere brain slices. Using python, images were thresholded with the Otsu threshold. Shape features were extracted from the binarized images. These shape features were analyzed based on brain region (cortex, hippocampus, thalamus), generalized treatment type (non-treated, injured, injured with treatment), and specific treatment type (OGD 0.5 hour, 1.5 hours, 3 hours, 1.5 hours with azithromycin treatment, 3 hours with superoxide dismutase treatment) and visualized using seaborn. The results verified trends in effects of injury and recovery after treatment on extent and solidity. Both findings support the expected shift from a circular shape of microglia in the injured state to more branched in the healthier state. The Otsu thresholding is limited in its accuracy, and, hence, these results provide an opportunity to optimize cell segmentation protocol for higher quality thresholded images. The results of this work have the potential to be applied to various forms of injury and cell types.