Comparison of unbiased estimation of neuronal number in the rat hippocampus with different staining methods

•The accuracy of neuronal number in hippocampus is particularly important in neuropathological processes and treatments.•The stereological method offers 3-D unbiased estimation for the total number of neurons in a defined brain region.•We provided a reliable and feasible method for quantification of total neuronal numbers by using unbiased stereological estimation.