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Abstract: The changes of directional diffusivities derived from diffusion tensor imaging (DTI), i.e. decreased axial diffusivity (λ ||) and increased radial diffusivity (λ ⊥), have shown significant correlation with axonal and myelin damage, respectively. However, after formalin fixation, reduced sensitivity of λ || in detecting axonal damage in tissue has raised the concern of applying DTI ex vivo. In order to distinguish whether death or the fixation process diminishes the sensitivity of DTI in detecting lesions, in vivo, pre-fixed postmortem, and fixed postmortem DTI were conducted on mouse optic nerves 3 and 14 days after transient retinal ischemia. Our data showed that, from in vivo to pre-fixed postmortem, λ || and λ ⊥ decreased by 50 to 70% in both healthy and injured optic nerves (3 and 14 day injury). From pre-fixed postmortem to fixed postmortem, λ || and λ ⊥ decreased by 40 to 50% in normal and 3-day injured optic nerves, but only by 15 to 25% in 14-day injured optic nerves. Consequently, for the 14-day injured optic nerves, the differences between healthy and injured nerves were not preserved after fixation: the 40% decreased λ|| and 200% increased λ⊥ in injured nerves as compared to the normal nerves were measured in vivo and pre-fixed postmortem, but after the fixation process, 300% increased λ⊥ and insignificant changes in λ|| were found in injured nerves as compared to the normal nerves. This study clarified that fixation process, but not death, could change the sensitivity of DTI in detecting injury. [Copyright &y& Elsevier]

Abstract: Decreased axial (λ ||) and increased radial (λ ⊥) diffusivity have been shown to reflect axonal and myelin injury respectively. In the present study, evolving white matter injury within the optic nerves of mice with retinal ischemia was examined by in vivo and ex vivo measurements of λ || and λ ⊥. The results show that at 3 days after retinal ischemia, a 33% decrease in vivo and a 38% decrease ex vivo in λ || without change in λ ⊥ was observed in the injured optic nerve compared to the control, suggestive of axonal damage without myelin injury. At 14 days, both in vivo and ex vivo measured λ ⊥ increased significantly to 220–240% of the control level in the injured optic nerve suggestive of myelin damage. In contrast, the axonal injury that was clearly detected in vivo as a significantly decreased λ || (33% decrease) was not as clearly detected by ex vivo λ || (17% decrease). The current findings suggest that ex vivo λ ⊥ is comparable to in vivo λ ⊥ in detecting myelin injury. However, the structural changes resulting from axonal damage causing the decreased in vivo λ || may not be preserved ex vivo in the fixed tissues. Despite the accurate depiction of the pathology using λ || and λ ⊥ in vivo, the use of ex vivo λ || to extrapolate the status of axonal injury in vivo would require further investigation. [Copyright &y& Elsevier]