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Image title	Accurate registration of bones from different developmental stages is achieved by matching the cortical cores.(A) A time chart of calcein and alizarin injection regimes. To cover the entire period effectively, three marginally overlapping regimes were applied. (B) Histological sections from each of the three injection regimes. At E18.5, green-labeled regions are seen around the middle part of the cortical core in both humerus and femur sections, indicating temporally preserved mineral structures. Similar temporal preservation of the cortical core is exhibited during perinatal and early postnatal development. (C) Color-coded statistical maps depicting the fraction of bones that are mineralized at each point in the bone, based on frontal slices of the femur. Clusters are composed of all bones from a single developmental day (top row), pairs (middle row), and triplets (bottom row) of consecutive developmental days. Regions that are highly preserved in population over several days (color-coded red; middle and bottom rows) are located mostly around the middle part of the cortical core. (D) A time series of frontal slices of the femur with the cortical core highlighted in each image with a different color. On the right is an overlay of all cortical core highlights following anatomical matching. The high overlap between segments demonstrates the potential of the cortical core to serve as a reliable salient structure for accurate anatomical matching of embryonic and early postnatal bones. Scale bars: 500μm in B–D, 25 μm in B insets I and II, and 50 μm in B inset III.
Copyright holder	Creative Commons Attribution License
Date(s)	4 August 2015
Publisher	Public Library of Science
Short title	Fig 3
Source	info:doi/10.1371/journal.pbio.1002212
Identifier	info:doi/info:doi/10.1371/journal.pbio.1002212.g003

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