ArticleNature protocols2021
Intravital microscopy of dynamic single-cell behavior in mouse mammary tissue.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 44 citations in OpenAlex.
- Quantitative rFRC-based evaluation of computational resolution enhancement in in vivo two-photon subcellular imaging.Biomedical optics express · 2026Article
- Loss of Cathepsin Z enhances pro-inflammatory macrophage responses and promotes tissue regeneration.Disease models & mechanisms · 2026Article
- Branching out: the diverse roles of stem and progenitor cells in mammary gland development.Journal of mammary gland biology and neoplasia · 2026Review
- BEHAV3D Tumor Profiler to map heterogeneous cancer cell behavior in the tumor microenvironment.eLife · 2025Article
- Next generation lineage tracing and its applications to unravel development.NPJ systems biology and applications · 2025Review
- Live imaging of the extracellular matrix with a glycan-binding fluorophore.Nature methods · 2025Article
- Noninvasive in vivo microscopy of single neutrophils in the mouse brain via NIR-II fluorescent nanomaterials.Nature protocols · 2024Review
- Imaging the extracellular matrix in live tissues and organisms with a glycan-binding fluorophore.bioRxiv : the preprint server for biology · 2024Article
- Cellular heterogeneity in TNF/TNFR1 signalling: live cell imaging of cell fate decisions in single cells.Cell death & disease · 2024Review
- Article
- Imaging actin organisation and dynamics in 3D.Journal of cell science · 2024Article
- A Decision Tree to Guide Human and Mouse Mammary Organoid Model Selection.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Recent advancements in single-cell metabolic analysis for pharmacological research.Journal of pharmaceutical analysis · 2023Review
- Spatially coordinated cell cycle activity and motility govern bifurcation of mammary branches.The Journal of cell biology · 2023Article
- Precise photoelectrochemical tuning of semiconductor microdisk lasers.Advanced photonics · 2023Article
- Multidimensional Imaging of Breast Cancer.Cold Spring Harbor perspectives in medicine · 2023Review
- Article
- Volume imaging to interrogate cancer cell-tumor microenvironment interactions in space and time.Frontiers in immunology · 2023Review
- Intravital imaging to study cancer progression and metastasis.Nature reviews. Cancer · 2023Review
- A Label-Free Segmentation Approach for Intravital Imaging of Mammary Tumor Microenvironment.Journal of visualized experiments : JoVE · 2022Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Multiphoton intravital imaging is essential for understanding cellular behavior and function in vivo. The adipose-rich environment of the mammary gland poses a unique challenge to in vivo microscopy due to light scattering that impedes high-resolution imaging. Here we provide a protocol for high-quality, six-color 3D intravital imaging of regions across the entire mouse mammary gland and associated tissues for several hours while maintaining tissue access for microdissection and labeling. An incision at the ventral midline and along the right hind leg creates a skin flap that is then secured to a raised platform skin side down. This allows for fluorescence-guided microdissection of connective tissue to provide unimpeded imaging of mammary ducts. A sealed imaging chamber over the skin flap creates a stable environment while maintaining access to large tissue regions for imaging with an upright microscope. We provide a strategy for imaging single cells and the tissue microenvironment utilizing multicolor Confetti lineage-tracing and additional dyes using custom-designed filters and sequential excitation with dual multiphoton lasers. Furthermore, we describe a strategy for simultaneous imaging and photomanipulation of single cells using the Olympus SIM scanner and provide steps for 3D video processing, visualization and high-dimensional analysis of single-cell behavior. We then provide steps for multiplexing intravital imaging with fixation, immunostaining, tissue clearing and 3D confocal imaging to associate cell behavior with protein expression. The skin-flap surgery and chamber preparation take 1.5 h, followed by up to 12 h of imaging. Applications range from basic filming in 1 d to 5 d for multiplexing and complex analysis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.