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Updated: May 13, 2026
Two-photon Imaging of Cellular Dynamics in the Mouse Spinal Cord
Published on: February 22, 2015
Two-photon imaging of immune cells in neural tissue
Insights
Observing immune cell function in the central nervous system (CNS) is key for developing new therapies. Two-photon microscopy visualizes immune cells in mouse brain slices and in vivo models of multiple sclerosis, aiding neuroinflammation research.
Area of Science:
- Neuroscience
- Immunology
- Microscopy
Background:
- Central nervous system (CNS) diseases require understanding immune cell behavior within neural tissue.
- Chronic neuroinflammation involves complex immune cell dynamics.
- Two-photon laser-scanning microscopy offers in vivo-like imaging of cellular functions.
Purpose of the Study:
- To detail a protocol for visualizing immune cell motility and function in CNS disease models.
- To provide methods for studying T helper 17 (Th17) cell migration in neuroinflammation.
- To enhance imaging techniques for CNS research.
Main Methods:
- Coculturing acute murine hippocampal slices with immune cells.
- In vivo imaging of the brain stem in experimental autoimmune encephalomyelitis (EAE) mouse models.
- Utilizing dual-color, two-photon laser-scanning microscopy for sample analysis.
Main Results:
- The protocol enables observation of T helper 17 (Th17) cell migration into brain parenchyma.
- Successful in vivo imaging of the brain stem in EAE mice was achieved.
- Technical aspects of microscopy allowing fluorophore flexibility were discussed.
Conclusions:
- This protocol facilitates the study of immune cell dynamics in CNS disease models.
- The methods support research into chronic neuroinflammation and potential therapeutic strategies.
- Advanced two-photon microscopy techniques improve visualization of neural and immune interactions.
Abstract:
To develop new therapeutic strategies for many central nervous system (CNS) diseases, it is essential to observe the motility and function of immune cells within neural tissue. Two-photon laser-scanning microscopy is an outstanding technique for imaging these phenomena under in vivo-like conditions. To gain deeper insight into the pathological phenomena that occur during chronic neuroinflammation of the CNS, we use it to view acute murine hippocampal slices cocultured with different subpopulations of immune cells and to view in vivo the brain stem of anesthetized transgenic mice affected by experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis. This protocol describes the preparation of cocultures of acute hippocampal slices with antigen-specific T helper 17 (Th17) cells migrating into the parenchyma, and the preparation of anesthetized mice for imaging the brain stem. We also discuss technical aspects of dual-color, two-photon laser-scanning microscopy that is used to image these samples and that allows for greater flexibility in the choice of fluorophores.
