ArticleBreast cancer research : BCR2016
Imaging the mammary gland and mammary tumours in 3D: optical tissue clearing and immunofluorescence methods.
Article in Breast cancer research : BCR, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 56 papers.
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56 citing papers in PubMed.
- Review
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- Little women: the relevance and reliance on mouse models for mammary gland research and next steps for translation.Journal of mammary gland biology and neoplasia · 2025Review
- Contractile fibroblasts form a transient niche for the branching mammary epithelium.Nature communications · 2025Article
- Retrograde tracing of breast cancer-associated sensory neurons.Journal of microscopy · 2025Article
- From morphology to single-cell molecules: high-resolution 3D histology in biomedicine.Molecular cancer · 2025Review
- Noncanonical Wnt/Ror2 Signaling Regulates Basal Cell Fidelity and Branching Morphogenesis in the Mammary Gland.bioRxiv : the preprint server for biology · 2025Article
- C/EBPβ deletion in macrophages impairs mammary gland alveolar budding during the estrous cycle.Life science alliance · 2024Article
- An ovine model for investigation of the microenvironment of the male mammary gland.Journal of anatomy · 2024Article
- Spatially distinct epithelial and mesenchymal cell subsets along progressive lineage restriction in the branching embryonic mammary gland.The EMBO journal · 2024Article
- A xenotransplantation mouse model to study physiology of the mammary gland from large mammals.PloS one · 2024Article
- Fibroblast-induced mammary epithelial branching depends on fibroblast contractility.PLoS biology · 2024Article
- Multidimensional Imaging of Breast Cancer.Cold Spring Harbor perspectives in medicine · 2023Review
- Article
- Quantifying Podocyte Number in a Small Sample Size of Glomeruli with CUBIC to Evaluate Podocyte Depletion of db/db Mice.Journal of diabetes research · 2023Article
- Three-dimensional imaging and analysis of pathological tissue samples with de novo generation of citrate-based fluorophores.Science advances · 2022Article
- Understanding Breast Cancers through Spatial and High-Resolution Visualization Using Imaging Technologies.Cancers · 2022Review
- Optical tissue clearing associated with 3D imaging: application in preclinical and clinical studies.Histochemistry and cell biology · 2022Review
- The immune environment of the mammary gland fluctuates during post-lactational regression and correlates with tumour growth rate.Development (Cambridge, England) · 2022Article
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7 authors.
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Abstract
backgroundHigh-resolution 3D imaging of intact tissue facilitates cellular and subcellular analyses of complex structures within their native environment. However, difficulties associated with immunolabelling and imaging fluorescent proteins deep within whole organs have restricted their applications to thin sections or processed tissue preparations, precluding comprehensive and rapid 3D visualisation. Several tissue clearing methods have been established to circumvent issues associated with depth of imaging in opaque specimens. The application of these techniques to study the elaborate architecture of the mouse mammary gland has yet to be investigated.
methodsMultiple tissue clearing methods were applied to intact virgin and lactating mammary glands, namely 3D imaging of solvent-cleared organs, see deep brain (seeDB), clear unobstructed brain imaging cocktails (CUBIC) and passive clarity technique. Using confocal, two-photon and light sheet microscopy, their compatibility with whole-mount immunofluorescent labelling and 3D imaging of mammary tissue was examined. In addition, their suitability for the analysis of mouse mammary tumours was also assessed.
resultsVarying degrees of optical transparency, tissue preservation and fluorescent signal conservation were observed between the different clearing methods. SeeDB and CUBIC protocols were considered superior for volumetric fluorescence imaging and whole-mount histochemical staining, respectively. Techniques were compatible with 3D imaging on a variety of platforms, enabling visualisation of mammary ductal and lobulo-alveolar structures at vastly improved depths in cleared tissue.
conclusionsThe utility of whole-organ tissue clearing protocols was assessed in the mouse mammary gland. Most methods utilised affordable and widely available reagents, and were compatible with standard confocal microscopy. These techniques enable high-resolution, 3D imaging and phenotyping of mammary cells and tumours in situ, and will significantly enhance our understanding of both normal and pathological mammary gland development.
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