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. 2023 Sep 23;4(4):102603. doi: 10.1016/j.xpro.2023.102603

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Whole-mount immunofluorescence staining following OMAR photobleaching

(A) The spatial distribution of three proteins (red channel): SOX9 (left panels), phospho-Histone H3 (pH 3, middle panels), and phospho-SMAD1/5/9 (pSMAD, right panels). Nuclei are counterstained by DAPI (blue channel).

(B) 3D reconstruction of the limb buds stained for SOX9 (left panels in A) and pSMAD (right panels in A) reveals the tissue integrity and detection of proteins throughout the limb bud with a very high signal-to-noise ratio. All limb buds shown are analyzed by confocal microscopy without digital post-processing apart from standard brightness and contrast adjustments. Scale bar: 500 μm.