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Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window


Holtmaat, A and Bonhoeffer, T and Chow, DK and Chuckowree, JA and De Paola, V and Hofer, SB and Hubener, M and Keck, T and Knott, G and Lee, WCA and Mostany, R and Mrsic-Flogel, TD and Nedivi, E and Portera-Cailliau, C and Svoboda, K and Trachtenberg, JT and Wilbrecht, L, Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window, Nature Protocols (Print), 4, (8) pp. 1128-1144. ISSN 1754-2189 (2009) [Refereed Article]

DOI: doi:10.1038/nprot.2009.89


To understand the cellular and circuit mechanisms of experience-dependent plasticity, neurons and their synapses need to be studied in the intact brain over extended periods of time. Two-photon excitation laser scanning microscopy (2PLSM), together with expression of fluorescent proteins, enables high-resolution imaging of neuronal structure in vivo. In this protocol we describe a chronic cranial window to obtain optical access to the mouse cerebral cortex for long-term imaging. A small bone flap is replaced with a coverglass, which is permanently sealed in place with dental acrylic, providing a clear imaging window with a large field of view (B0.8–12 mm2). The surgical procedure can be completed within B1 h. The preparation allows imaging over time periods of months with arbitrary imaging intervals. The large size of the imaging window facilitates imaging of ongoing structural plasticity of small neuronal structures in mice, with low densities of labeled neurons. The entire dendritic and axonal arbor of individual neurons can be reconstructed.

Item Details

Item Type:Refereed Article
Research Division:Biomedical and Clinical Sciences
Research Group:Neurosciences
Research Field:Neurosciences not elsewhere classified
Objective Division:Health
Objective Group:Clinical health
Objective Field:Clinical health not elsewhere classified
UTAS Author:Chuckowree, JA (Dr Jyoti Chuckowree)
ID Code:62287
Year Published:2009
Web of Science® Times Cited:618
Deposited By:Menzies Institute for Medical Research
Deposited On:2010-03-11
Last Modified:2010-04-15

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