In vivo multiphoton nanosurgery on cortical neurons

Year: 2007

Authors: Sacconi L., O’Connor R.P., Jasaitis A., Masi A., Buffelli M., Pavone FS.

Autors Affiliation: University of Florence, European Laboratory for Nonlinear Spectroscopy, Florence, Italy
University of Florence, Department of Physics, Florence, Italy
University of Florence, Department of Pathology and Experimental Oncology, Florence, Italy
University of Verona, Department of Neurological and Visual Sciences, Verona, Italy

Abstract: Two-photon microscopy has been used to perform high spatial resolution imaging of spine plasticity in the intact neocortex of living mice. Multiphoton absorption has also been used as a tool for the selective disruption of cellular structures in living cells and simple organisms. In this work, we exploit the spatial localization of multiphoton excitation to perform selective lesions on the neuronal processes of cortical neurons in living mice expressing fluorescent proteins. Neurons are irradiated with a focused, controlled dose of femtosecond laser energy delivered through cranial optical windows. The morphological consequences are then characterized with time lapse 3-D two-photon imaging over a period of minutes to days after the procedure. This methodology is applied to dissect single dendrites with submicrometric precision without causing any visible collateral damage to the surrounding neuronal structures. The spatial precision of this method is demonstrated by ablating individual dendritic spines, while sparing the adjacent spines and the structural integrity of the dendrite. The combination of multiphoton nanosurgery and in vivo imaging in mammals represents a promising tool for neurobiology and neuropharmacology research. (c) 2007 Society of Photo-Optical Instrumentation Engineers.


Volume: 12 (5)      Pages from: 050502-1  to: 050502-3

KeyWords: in vivo imaging; long-term imaging; two-photon microscopy; laser ablation; laser surgery
DOI: 10.1117/1.2798723

Citations: 59
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