Neuroimaging studies have routinely used hippocampal volume as a measure of Alzheimer’s disease severity, but hippocampal changes occur too late in the disease process for potential therapies to be effective. The entorhinal cortex is one of the first cortical areas affected by Alzheimer’s disease; its neurons are especially vulnerable to neurofibrillary tangles. Entorhinal atrophy also relates to the conversion from nonclinical to clinical Alzheimer’s disease. In neuroimaging, the human entorhinal cortex has so far mostly been considered in its entirety or divided into a medial and a lateral region. Cytoarchitectonic differences provide the opportunity for subfield parcellation. We investigated the entorhinal cortex on a subfield-specific level – at a critical time point of Alzheimer’s disease progression. While magnet resonance imaging allows multidimensional quantitative measurements, only histology provides enough accuracy to determine subfield boundaries - the prerequisite for quantitative measurements within the entorhinal cortex. This study used histological data to validate ultra-high resolution 7 Tesla ex vivo MRI and create entorhinal subfield parcellations in a total of 10 preclinical Alzheimer’s disease and normal control cases. Using ex vivo magnet resonance imaging eight entorhinal subfields (olfactory, rostral, medial intermediate, intermediate, lateral rostral, lateral caudal, caudal, caudal limiting) were characterized for cortical thickness, volume, and pial surface area. Our data indicated no influence of sex, or Braak and Braak staging on volume, cortical thickness, or pial surface area. The volume and pial surface area for mean whole entorhinal cortex was 1131 mm3 ± 55.72 and 429 mm2 ± 22.6 (mean ± S.E.M), respectively. The subfield volume percentages relative to the entire entorhinal cortex were: olfactory: 18.73% ± 1.82, rostral: 14.06% ± 0.63, lateral rostral: 14.81% ± 1.22, medial intermediate: 6.72% ± 0.72, intermediate: 23.36% ± 1.85, lateral caudal: 5.42% ± 0.33, caudal: 10.99% ± 1.02, and caudal limiting: 5.91% ± 0.40 (all mean ± standard error of the mean). Olfactory and intermediate subfield revealed the most extensive intra-individual variability (cross-subject variance) in volume and pial surface area. This study provides validated measures. It maps individuality and demonstrates human variability in the entorhinal cortex, providing a baseline for approaches in individualized medicine. Taken together, this study serves as a ground truth validation study for future in vivo comparisons and treatments.