This study introduces a methodology for generating high resolution signal profiles of microcalcification (MC) grains for validating breast CT (bCT) systems. A physical MC phantom was constructed by suspending calcium carbonate grains in an agar solution emulating MCs in a fibroglandular tissue background. Additionally, small Teflon spheres (2.4 mm diameter) were embedded in the agar solution for the purpose of fiducial marking and assessment of segmentation accuracy. The MC phantom was imaged on a high resolution (34 µm) commercial small-bore µCT scanner at high dose, and the images were used as the gold-standard for assessing MC size and for generating high resolution signal profiles of each MC. High-dose bCT scans of the MC phantom suspended in-air were acquired using 1×1 binning mode (75 µm dexel pitch) by averaging three repeat scans to produce a single low-noise reconstruction of the MC phantom. The high resolution µCT volume data set was then registered with the corresponding bCT data set after correcting for the bCT system spatial resolution. Microcalcification signal profiles constructed using low-noise bCT images were found to be in good agreement with those generated using the µCT scanner with all differences < 10% within the VOI surrounding each MC. The MC signal profiles were used as detection templates for a non-prewhitening-matched-filter model observer for scans acquired in a realistic breast phantom at 3, 6, and 9 mGy mean glandular dose. MC detectability using signal templates derived from bCT were shown to be in good agreement with those generated using µCT.