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Fig. 4 | Breast Cancer Research

Fig. 4

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

Fig. 4

Clear unobstructed brain imaging cocktails (CUBIC) clearing for whole-mount transmission imaging of the mouse mammary gland. a Virgin and lactating mammary glands stained with methyl green and cleared with CUBIC for whole-mount morphometric analysis. These images are representative of images from more than three mice. b Methyl green counterstaining, showing the compatibility of this light green counterstain with magenta-glu detection of β-glucosidase+ cells; β-glucosidase+ cells are interspersed with unlabelled cells in this R26 [CA]30SYNbglA mouse model. c Compatibility of CUBIC clearing with smooth muscle actin (SMA)-immunostaining and horseradish peroxidase-3,3-diaminobenzidine detection. Immunostaining steps can be performed before CUBIC clearing (top panel) or after CUBIC clearing (bottom panel). 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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