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Chronic Imaging of Mouse Visual Cortex Using a Thinned-skull Preparation
Published on: October 25, 2010
Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.
Anthony Holtmaat1, Tobias Bonhoeffer, David K Chow
1Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, NY, USA. anthony.holtmaat@medecine.unige.ch
Nature Protocols
|July 21, 2009
Summary
This study details a chronic cranial window technique for long-term in vivo imaging of neuronal plasticity in mice. This method allows detailed observation of cellular and circuit mechanisms over months.
Area of Science:
- Neuroscience
- Cell Biology
- Microscopy
Background:
- Understanding experience-dependent plasticity requires long-term study of neurons in vivo.
- High-resolution imaging of neuronal structure in vivo is crucial.
Purpose of the Study:
- To describe a chronic cranial window protocol for long-term optical imaging of the mouse cerebral cortex.
- To enable detailed study of neuronal structure and plasticity over extended periods.
Main Methods:
- A chronic cranial window is surgically created by replacing a bone flap with a sealed coverglass.
- Two-photon excitation laser scanning microscopy (2PLSM) is used for high-resolution imaging.
- The protocol allows for imaging over months with flexible intervals.
Main Results:
- The chronic cranial window provides a large field of view (approx. 0.8-12 mm(2)) for in vivo imaging.
- The preparation supports imaging of structural plasticity in small neuronal structures.
- Individual neurons' dendritic and axonal arbors can be fully reconstructed.
Conclusions:
- The chronic cranial window is a valuable tool for long-term in vivo neuroscience research.
- This technique facilitates the study of experience-dependent plasticity at cellular and circuit levels.
- The method allows for detailed reconstruction of neuronal morphology over time.
