Development of efficient imaging techniques to trace neuronal connections would be very useful. Manganese ion (Mn2+) is an excellent T1 contrast agent for magnetic resonance imaging (MRI). Four reports utilizing radioactive Mn2+ in fish and rat brain indicate that Mn2+ may be useful for tracing neuronal connections. Therefore, the purpose of this work was to determine if Mn2+ can be used as an in vivo MRI neuronal tract tracer. The results indicate that topical administration of MnCI2 solution to the naris of mice as well as to the retinal ganglion cells via intravitreal injection leads to enhancement of contrast along the respective pathways. Therefore, application of Mn2+ to neurons allows the use of MRI to visualize neuronal connections.