Dendra2 photoswitching through the Mammary Imaging Window

Area of Science:

• Oncology
• Microscopy
• Animal Models

Background:

• Intravital microscopy provides critical insights into tumor metastasis and immune responses.
• Previous methods limited long-term imaging of orthotopic breast cancer models.
• Understanding tumor microenvironments is key to developing effective cancer therapies.

Purpose of the Study:

• To describe a protocol for manufacturing and utilizing a Mammary Imaging Window (MIW).
• To enable long-term, single-cell resolution intravital imaging of orthotopic mammary carcinomas in mice.
• To facilitate the study of tumor cell migration, angiogenesis, and host cell interactions in vivo.

Main Methods:

• Development of a Mammary Imaging Window (MIW).
• Optimization of Dendra2 photoswitching and in vivo imaging parameters.
• Detailed protocol for MIW insertion and imaging of Dendra2-labeled tumor cells using a custom imaging box.

Main Results:

• The MIW protocol allows for repeated intravital imaging sessions in living mice.
• High-resolution imaging of Dendra2-labeled tumor cells, blood vessels, and host cells is achievable.
• The technique enables visualization of tumor cell behavior within distinct microenvironments.

Conclusions:

• The described protocol offers a robust method for long-term intravital imaging of breast cancer.
• This technique advances the study of metastatic mechanisms and tumor microenvironment dynamics.
• It provides a valuable tool for preclinical research in breast cancer metastasis and therapy evaluation.