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. 2016 Dec 13;18:127. doi: 10.1186/s13058-016-0754-9

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© The Author(s). 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 2 — Passive clarity technique (PACT)-sorbitol refractive index matching solution (sRIMS) clearing and 3D imaging of virgin and lactating mouse mammary tissue. a PACT-sRIMS tissue clearing and immunostaining protocol and timeline. Three-dimensional confocal imaging of PACT-sRIMS-cleared virgin (b) and lactating (c) mammary glands immunostained with basal cell markers (K5 and smooth muscle actin (SMA)) and luminal cell markers (K8 and E-cadherin (E-CAD)). Main image shows the maximum intensity projection of the entire image sequence, with thin optical slices (1 μm) and their depth (z value) relative to the first image in the image sequence. These images are representative of images from at least two mice; further examples of PACT-sRIMS-cleared tissue are shown in Additional file 6: Figure S6). DAPI 4′,6-diamidino-2-phenylindole. See additional file 18 for a high resolution version of these PDFs and http://rdcu.be/lT3Z for additional high resolution examples of these imaging techniques [38]