Rationale and Objectives: Breast density is a significant breast cancer risk factor measured from mammograms. Evidence suggests that the spatial variation in mammograms may also be associated with risk. We investigated the variation in calibrated mammograms as a breast cancer risk factor and explored its relationship with other measures of breast density using full field digital mammography (FFDM). Materials and Methods: A matched case-control analysis was used to assess a spatial variation breast density measure in calibrated FFDM images, normalized for the image acquisition technique variation. Three measures of breast density were compared between cases and controls: (a) the calibrated average measure, (b) the calibrated variation measure, and (c) the standard percentage of breast density (PD) measure derived from operator-assisted labeling. Linear correlation and statistical relationships between these three breast density measures were also investigated. Results: Risk estimates associated with the lowest to highest quartiles for the calibrated variation measure were greater in magnitude (odds ratios: 1.0 [ref.], 3.5, 6.3, and 11.3) than the corresponding risk estimates for quartiles of the standard PD measure (odds ratios: 1.0 [ref.], 2.3, 5.6, and 6.5) and the calibrated average measure (odds ratios: 1.0 [ref.], 2.4, 2.3, and 4.4). The three breast density measures were highly correlated, showed an inverse relationship with breast area, and related by a mixed distribution relationship. Conclusion: The three measures of breast density capture different attributes of the same data field. These preliminary findings indicate the variation measure is a viable automated method for assessing breast density. Insights gained by this work may be used to develop a standard for measuring breast density.