Intravital Microscopy for Imaging the Tumor Microenvironment in Live Mice

  • Victor Naumenko
  • Craig JenneEmail author
  • Douglas J. MahoneyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1458)


The development of intravital microscopy has provided unprecedented capacity to study the tumor microenvironment in live mice. The dynamic behavior of cancer, stromal, vascular, and immune cells can be monitored in real time, in situ, in both primary tumors and metastatic lesions, allowing treatment responses to be observed at single cell resolution and therapies tracked in vivo. These features provide a unique opportunity to elucidate the cellular mechanisms underlying the biology and treatment of cancer. We describe here a method for imaging the microenvironment of subcutaneous tumors grown in mice using intravital microscopy.

Key words

Tumor microenvironment Intravital microscopy Multiphoton imaging Confocal imaging Mouse models Subcutaneous tumors 

Supplementary material

Supplemental Video 1

4D multiphoton imaging of tumor microenvironment. Multiphoton intravital imaging of tumor cells (red), vessels (gray), virus infection (green), collagen/second harmonic (cyan). Image is composed of a z-stack of ten optical sections (8 μm per section) (MP4 2372 kb)

Supplemental Video 2

Spinning-disk imaging of lymphocyte populations within tumor vasculature. Spinning-disk confocal intravital imaging of CD4 (green), CD8 (blue) and NK cells (red) within the vasculature of a B16 tumor (MP4 761 kb)

Supplemental Video 3

Resonant-scanning confocal imaging of leukocytes within the tumor vasculature. Intravital imaging of cells stained for CD11b (red) and Ly6G (green) stained cells within vasculature or a CT26 tumor (MP4 3930 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Alberta Children’s Hospital Research InstituteCalgaryCanada
  2. 2.Arnie Charbonneau Cancer InstituteCalgaryCanada
  3. 3.Snyder Institute for Chronic DiseasesCalgaryCanada
  4. 4.Department of Microbiology, Immunology and Infectious Diseases, Faculty of MedicineUniversity of CalgaryCalgaryCanada

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