Autopsy studies of Alzheimer's disease (AD) have found that neurofibrillary tangle (NFT) pathology of the medial temporal lobe (MTL) demonstrates selective topography with relatively stereotyped subregional involvement at early disease stages, prompting interest in more granular measurement of these structures with in vivo magnetic resonance imaging. We applied a novel, automated method for measurement of hippocampal subfields and extrahippocampal MTL cortical regions. The cohort included cognitively normal (CN) adults (n = 86), early mild cognitive impairment (n = 43), late MCI (n = 22), and mild AD (n = 40) patients from the Alzheimer's Disease Neuroimaging Initiative (ADNI). For pseudolongitudinal analysis of the continuum from preclinical to mild AD dementia, the groups were further divided according to amyloid status based on positron emission tomography. Specific subregions associated with the early NFT pathology of AD were more sensitive to preclinical and early prodromal AD than whole hippocampal volume while more diffuse involvement was found in later stages. In particular, BA35, the first region associated with NFT deposition, was the only region to discriminate preclinical AD from amyloid negative cognitively normal adults ("normal aging"). In general, patterns of atrophy in the pseudolongitudinal analysis largely recapitulated Braak staging of NFTs within the MTL.