Optical coherence tomography (OCT) is widely used to noninvasively assess retinal nerve fiber layer (RNFL) thickness to assess optic nerve damage. However, the impact of coexisting pathology on OCT-detected RNFL thickness remains unclear. We have previously developed a diffusion basis spectrum imaging (DBSI) to directly assess coexisting pathologies in mouse models of optic nerve crush (ONC) and optic neuritis. In the current study, we assessed retinal pathology and coexisting pathologies in optic nerve of ONC mice longitudinally, with OCT and DBSI, respectively. Our goal is to determine the relationship between OCT-detected RNFL change and DBSI-derived optic nerve pathologies.