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Updated: May 22, 2026
A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation
Published on: September 19, 2010
Optical window preparation for two-photon imaging of microglia in mice
Abstract:
Microglia are the primary immune effector cells of the brain parenchyma. They are distributed throughout the brain at various densities. Two-photon fluorescence microscopy, together with expression of fluorescent proteins in microglia, has enabled the study of these fascinating cells in vivo. Imaging studies have shown, for example, that microglia continually survey their cellular environment and immediately respond to injury. However, we still know very little about their roles in various parts of the developing and adult brain or their diverse effector functions in aging and different disease states. Experimental procedures have been developed for minimally invasive short- and long-term two-photon imaging of microglial cells in cortical regions of the intact mouse brain. This protocol presents two methods for the preparation of the optical window that is needed for two-photon imaging of microglia. The thinned skull method should be used whenever possible. Skull thinning enables transcranial two-photon imaging while minimizing external influences that might disturb normal microglia physiology and brain homeostasis. The sealed craniotomy preparation is useful for short-term investigation of microglia.
Insights
This study details methods for in vivo two-photon imaging of microglia, the brain's immune cells. These techniques allow researchers to observe microglia's dynamic behavior and responses in the intact mouse brain.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are the brain's primary immune cells, surveying and responding to their environment.
- Current understanding of microglial roles in development, aging, and disease remains limited.
- In vivo imaging techniques are crucial for studying microglia's dynamic functions.
Purpose of the Study:
- To present protocols for minimally invasive two-photon imaging of microglia in the mouse brain.
- To enable detailed observation of microglial behavior and physiology in vivo.
- To facilitate research into microglial functions across different brain regions and conditions.
Main Methods:
- Development of two distinct optical window preparation methods for transcranial two-photon microscopy.
- Skull thinning technique for long-term, minimally invasive imaging, preserving brain homeostasis.
- Sealed craniotomy preparation for short-term microglial investigations.
Main Results:
- Established protocols for high-resolution, in vivo imaging of microglia in the intact mouse cortex.
- Demonstrated the feasibility of observing microglial environmental surveying and injury responses.
- Provided methods to minimize external disturbances during imaging.
Conclusions:
- The presented methods enhance the ability to study microglia in their native environment.
- These techniques are vital for advancing our understanding of microglial roles in brain health and disease.
- Further research into microglial functions can be significantly aided by these imaging protocols.
