Precise Brain Mapping to Perform Repetitive In Vivo Imaging of Neuro-Immune Dynamics in Mice

Kanchan Bisht; Kaushik Sharma; Ukpong  B. Eyo

doi:10.3791/61454

Precise Brain Mapping to Perform Repetitive In Vivo Imaging of Neuro-Immune Dynamics in Mice

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Precise Brain Mapping to Perform Repetitive In Vivo Imaging of Neuro-Immune Dynamics in Mice

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08:17 min

August 7, 2020

DOI: 10.3791/61454-v

Kanchan Bisht^1,2, Kaushik Sharma^1,2, Ukpong B. Eyo^1,2

¹Center for Brain Immunology and Glia (BIG),University of Virginia, ²Department of Neuroscience,University of Virginia

Overview

This protocol demonstrates a chronic cranial window implantation technique for longitudinal imaging of neuro-glio-vascular structures in the brain. It allows for repetitive visualization of cellular dynamics using In Vivo Two-Photon microscopy, providing insights into cellular interactions and changes over time.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Imaging Techniques

Background

Longitudinal imaging is crucial for studying brain dynamics.
Transcranial imaging has limitations that this technique addresses.
Understanding cellular interactions is vital for insights into brain function.
Two-Photon microscopy enables detailed imaging of cellular elements.

Purpose of Study

To visualize long-term changes in brain cellular elements.
To study cell-cell interactions and dynamics during brain plasticity.
To investigate morphological changes in neurodegeneration.

Methods Used

Preparation of CX3CR1 GFP heterozygote mice for cranial window implantation.
Anesthesia and stabilization of the mouse during surgery.
Use of Two-Photon microscopy for imaging cellular dynamics.
Longitudinal studies of neuro-glio-vascular interactions.

Main Results

Successful implantation of cranial windows for imaging.
Visualization of cellular dynamics over extended periods.
Insights into the arrangement and morphology of CNS cells.
Identification of changes during brain plasticity and neurodegeneration.

Conclusions

The cranial window technique is effective for longitudinal studies.
Two-Photon microscopy provides valuable data on cellular interactions.
This approach enhances understanding of brain function in health and disease.

Frequently Asked Questions

What is the purpose of the cranial window implantation?

The cranial window allows for longitudinal imaging of brain structures and cellular dynamics.

How does Two-Photon microscopy benefit this study?

It enables repetitive visualization of cellular changes in the same brain location over time.

What type of mice are used in this protocol?

Juvenile or young adult CX3CR1 GFP heterozygote mice are used.

What are the main applications of this technique?

It is useful for studying brain plasticity, neurodegeneration, and cellular interactions.

What are the limitations of transcranial imaging?

Transcranial imaging has critical limitations that this protocol aims to address.

What preparations are needed before surgery?

The mouse must be anesthetized and its head shaved for proper implantation.

This protocol describes a chronic cranial window implantation technique that can be used for longitudinal imaging of neuro-glio-vascular structures, interactions, and function in both healthy and diseased conditions. It serves as a complementary alternative to the transcranial imaging approach that, while often preferred, possesses some critical limitations.

This protocol demonstrates how In Vivo Two-Photon microscopy can be used to do repetitive visualization of cellular dynamics, in the same brain location over extended time periods. The advantage of this technique is that, it makes it possible to image the longterm changes in cellular elements of brain, including the arrangement, morphology, and physical interactions between different cell type of the CNS. This technique can be especially useful in eliciting cell-cell interactions, cell dynamics, and morphological changes during brain plasticity, and neurodegeneration.

Begin by preparing a juvenile or young adult, four to 10 week old, a CX3CR1 GFP heterozygote mouse, that weighs 17 to 25 grams, for a cranial window implantation. After anesthetizing the mouse, use a hair trimmer to shave the hair on its head, between the ears, approximately, from the eye level, to the top of the neck region. Move the mouse to the stereotactic surgery station, nose cone, and stabilize its head using ear bars.

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