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Fig. 5 | Breast Cancer Research

Fig. 5

From: Imaging the mammary gland and mammary tumours in 3D: optical tissue clearing and immunofluorescence methods

Fig. 5

See deep brain (SeeDB)-clearing and 3D imaging of virgin and lactating mouse mammary tissue. a SeeDB tissue clearing and immunostaining protocol and timeline. Three-dimensional confocal imaging of SeeDB-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 (green) 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. Right panel shows depth-coding of SMA-expressing cells; images in an image stack are assigned a colour based on their relative depth. These images are representative of images from more than three mice; further examples of SeeDB-cleared tissue are shown in Additional file 13: Figure S11). BV blood vessel (SMA-expressing). 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]

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