title: Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window.
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.
authors.affiliation: Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, NY, USA. anthony.holtmaat@medecine.unige.ch
authors.forename: Anthony
authors.initials: A
authors.lastname: Holtmaat
authors.forename: Tobias
authors.initials: T
authors.lastname: Bonhoeffer
authors.forename: David K
authors.initials: DK
authors.lastname: Chow
authors.forename: Jyoti
authors.initials: J
authors.lastname: Chuckowree
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Paola
authors.forename: Sonja B
authors.initials: SB
authors.lastname: Hofer
authors.forename: Mark
authors.initials: M
authors.lastname: Hübener
authors.forename: Tara
authors.initials: T
authors.lastname: Keck
authors.forename: Graham
authors.initials: G
authors.lastname: Knott
authors.forename: Wei-Chung A
authors.initials: WC
authors.lastname: Lee
authors.forename: Ricardo
authors.initials: R
authors.lastname: Mostany
authors.forename: Tom D
authors.initials: TD
authors.lastname: Mrsic-Flogel
authors.forename: Elly
authors.initials: E
authors.lastname: Nedivi
authors.forename: Carlos
authors.initials: C
authors.lastname: Portera-Cailliau
authors.forename: Karel
authors.initials: K
authors.lastname: Svoboda
authors.forename: Joshua T
authors.initials: JT
authors.lastname: Trachtenberg
authors.forename: Linda
authors.initials: L
authors.lastname: Wilbrecht
mesh_terms: D000818:Animals; D003399:Craniotomy / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis; D051379:Mice; D008856:Microscopy, Fluorescence; D019579:Neocortex / Q000166:cytology* / Q000502:physiology; D009473:Neuronal Plasticity; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D012886:Skull / Q000601:surgery
chemical_list: D008164:Luminescent Proteins