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

Holtmaat, A., Bonhoeffer, T., Chow, D. K., Chuckowree, J., De Paola, V., Hofer, S. B., Hubener, M., Keck, T., Knott, G., Lee, W. C., Mostany, R., Mrsic-Flogel, T. D., Nedivi, E., Portera-Cailliau, C., Svoboda, K., Trachtenberg, J. T., Wilbrecht, L. (July 2009) Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window. Nat Protoc, 4 (8). pp. 1128-44. ISSN 1750-2799 (Electronic)

URL: http://www.ncbi.nlm.nih.gov/pubmed/19617885
DOI: 10.1038/nprot.2009.89

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.

Item Type: Paper
Uncontrolled Keywords: imaging model organisms neuroscience Craniotomy/ methods Animals Luminescent Proteins/analysis Mice Microscopy, Fluorescence Neocortex/ cytology/physiology Neuronal Plasticity Neurons/cytology/metabolism Skull/surgery
Subjects: Investigative techniques and equipment
organism description > animal
organs, tissues, organelles, cell types and functions > organs types and functions > brain
Investigative techniques and equipment > imaging
organism description > animal > mammal > rodent > mouse
organs, tissues, organelles, cell types and functions > organs types and functions
organs, tissues, organelles, cell types and functions
Investigative techniques and equipment > microscopy > flourescence microscopy > two-photon excitation microscopy
CSHL Authors:
Communities: CSHL labs > Svoboda lab
Depositing User: Matt Covey
Date: 16 July 2009
Date Deposited: 20 Feb 2013 19:29
Last Modified: 20 Feb 2013 19:29
PMCID: PMC3072839
Related URLs:
URI: http://repository.cshl.edu/id/eprint/27407

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