To study the effects of overlapping anatomy on microcalcification detection at various incident exposure levels. Images of an anthropomorphic breast phantom (RMI 169) overlapping with simulated microcalcifications ranging from 150 to 212 μn in size placed in two breast density regions, fatty and heterogencously dense, were acquired with an a-Si/a-Se flat panel based digital mammography system (Selenia) operated with Mo-Mo target/filter combination at 28 kVp. The mammograms were exposed with 20, 30, 40, 60, 80, 120, 160, 240 and 325 mAs for varying the exposure level. A 4-AFC study was performed for evaluation of the detection performance. Four 400×400-pixel images were displayed as 2×2 array on a LCD flat panel based review workstation. One of the four images contained a cluster of five microcalcifications and was randomly placed in one of the four quadrants. A physicist was asked to select the image containing the microcalcifications and to report the number of visible microcalcifications. The fraction of correct responses was computed with two different criteria: (1) the selected images contained one or more microcalcifications, and (2) the selected images contained 4 or 5 visible microcalcifications. The statistical significance of the differences in fractions for different exposure levels and regions was evaluated. The results showed that, if visibility of one or more microcalcifications is required, the fractions of correct responses were 1 for all size groups and most exposure levels in both fatty and heterogencously dense regions. If a visibility of 80% or more of the microcalcifications was required, the fractions of correct responses significantly decreased in both regions. The results indicated that microcalcification detection in the fatty region appeared to be mainly limited by the quantum noise, and that in the heterogencously dense region may be limited by both the anatomic noise and the quantum noise.