A customized affordable multiplexed immunofluorescence method visualizes early changes in the mouse brain...

Area of Science:

• Immunology
• Neuroscience
• Biotechnology

Background:

• Multiplex immunofluorescence (mIF) allows simultaneous visualization of multiple protein targets in tissue sections.
• Laser interstitial thermal therapy (LITT) is a minimally invasive treatment for brain tumors, causing tissue damage and potential immunomodulation.
• Understanding the immune response in the brain tumor microenvironment (TME) after LITT is crucial.

Purpose of the Study:

• To develop a versatile, affordable, and customizable mIF method for spatial imaging of early tissue responses in post-LITT mouse brains.
• To assess TME changes in formalin-fixed paraffin-embedded (FFPE) mouse brain tissue sections after LITT.
• To investigate the immunomodulatory roles of LITT in the brain TME.

Main Methods:

• Developed a customizable mIF protocol using standard histology and microscopy equipment.
• Combined mIF with a laser cytoreduction workflow monitored by MRI.
• Analyzed spatial distribution of multiple cellular and molecular immunotargets in FFPE mouse brain sections post-LITT.

Main Results:

• Enabled spatial visualization of resident brain cell activation states post-LITT.
• Showcased the emergence and distribution of diverse phagocytic immune cell populations at the post-LITT site.
• Demonstrated mIF's capability to analyze immune cell infiltration and resident brain cell responses in mouse brain TME.

Conclusions:

• Multiplex IF on mouse models advances understanding of repair and immune responses post-LITT.
• The customizable mIF protocol is cost-effective for investigating spatial distribution of immune cells and resident brain cell activation.
• This method aids in comprehending the complex interplay within the brain TME following LITT.