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

Holtmaat, Anthony; Bonhoeffer, Tobias; Chow, David K.; Chuckowree, Jyoti; De Paola, Vincenzo; Hofer, Sonja B.; Hübener, Mark; Keck, Tara; Knott, Graham; Lee, Wei-Chung A.; Mostany, Ricardo; Mrsic-Flogel, Tom D.; Nedivi, Elly; Portera-Cailliau, Carlos; Svoboda, Karel; Trachtenberg, Joshua T.; Wilbrecht, Linda

doi:10.1038/nprot.2009.89

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Title		Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window
Authors		Holtmaat, Anthony Bonhoeffer, Tobias Chow, David K. Chuckowree, Jyoti De Paola, Vincenzo Hofer, Sonja B. Hübener, Mark Keck, Tara Knott, Graham Lee, Wei-Chung A. Mostany, Ricardo Mrsic-Flogel, Tom D. Nedivi, Elly Portera-Cailliau, Carlos Svoboda, Karel Trachtenberg, Joshua T. Wilbrecht, Linda show all authors [1 - 17]
Published in		Nature Protocols. 2009, vol. 4, no. 8, p. 1128-1144
Abstract		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 (approximately 0.8-12 mm(2)). The surgical procedure can be completed within approximately 1 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.
Keywords		Animals — Craniotomy/methods — Luminescent Proteins/analysis — Mice — Microscopy, Fluorescence — Neocortex/cytology/physiology — Neuronal Plasticity — Neurons/cytology/metabolism — Skull/surgery
Identifiers		DOI: 10.1038/nprot.2009.89 PMID: 19617885
Full text		Article (Published version) (1.8 MB) - Limited access to UNIGE Other version: http://www.nature.com/nprot/journal/v4/n8/full/nprot.2009.89.html
Structures		Faculté de médecine / Section de médecine fondamentale / Département de neurosciences fondamentales
Research group		Les circuits neuronaux dans le néocortex des mammifères (855)
Citation (ISO format)		HOLTMAAT, Anthony et al. Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window. In: Nature Protocols, 2009, vol. 4, n° 8, p. 1128-1144. doi: 10.1038/nprot.2009.89 https://archive-ouverte.unige.ch/unige:5329