Deep intravital brain tumor imaging enabled by tailored three-photon microscopy and analysis

Area of Science:

• Neuroscience
• Oncology
• Medical Imaging

Background:

• Intravital 2P-microscopy is limited to superficial brain layers.
• Studying white matter invasion in glioblastoma has been challenging due to technical limitations.

Purpose of the Study:

• To develop a method for deep intravital imaging of glioblastoma in mouse white matter.
• To investigate glioblastoma invasion routes and mechanisms in the corpus callosum.
• To identify potential imaging biomarkers for early glioblastoma colonization.

Main Methods:

• Developed an AI-enhanced intravital microscopy workflow (Deep3P).
• Enabled longitudinal imaging of glioblastoma up to 1.2 mm depth in mouse brain.
• Analyzed tumor cell migration and morphological changes in white matter.

Main Results:

• Deep3P successfully imaged glioblastoma at unprecedented depths.
• Identified perivascular invasion as a primary route into the corpus callosum.
• Uncovered two distinct vascular mechanisms of glioblastoma migration.
• Observed morphological changes indicative of early colonization.

Conclusions:

• Deep3P facilitates non-invasive, deep intravital studies of brain tumors.
• Provides novel insights into glioblastoma invasion and the tumor microenvironment.
• Opens avenues for studying brain tumor neuroscience and developing imaging biomarkers.