In vivo multi-photon nanosurgery on cortical neurons: focusing on network organization

L. Sacconi; R. P. O'Connor; A. Jasaitis; A. Masi; M. Buffelli; F. S. Pavone

doi:10.1117/12.761588

15 February 2008 In vivo multi-photon nanosurgery on cortical neurons: focusing on network organization

L. Sacconi, R. P. O'Connor, A. Jasaitis, A. Masi, M. Buffelli, F. S. Pavone

Proceedings Volume 6860, Multiphoton Microscopy in the Biomedical Sciences VIII; 686024 (2008) https://doi.org/10.1117/12.761588
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

Two-photon microscopy has been used to perform high spatial resolution imaging of spine plasticity in the intact neocortex of living mice. Multi-photon 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 multi-photon excitation to perform selective lesions on the neuronal processes of cortical neurons in living mice expressing fluorescent proteins. This methodology was applied to dissect single dendrites with sub-micrometric precision without causing any visible collateral damage to the surrounding neuronal structures. The spatial precision of this method was demonstrated by ablating individual dendritic spines, while sparing the adjacent spines and the structural integrity of the dendrite. The morphological consequences were then characterized with time lapse 3D two-photon imaging over a period of minutes to days after the procedure. Here we present the results of our systematic study of the morphological response of cortical pyramidal neurons to nanosurgical perturbations. Dendritic branches were followed after transecting distal segments, whilst the plasticity and remodeling of individual dendritic spines on a given branch was also followed after removing of a subset of spines.

Citation Download Citation

L. Sacconi, R. P. O'Connor, A. Jasaitis, A. Masi, M. Buffelli, and F. S. Pavone "In vivo multi-photon nanosurgery on cortical neurons: focusing on network organization", Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 686024 (15 February 2008); https://doi.org/10.1117/12.761588

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KEYWORDS

Dendrites

Spine

Neurons

In vivo imaging

Fluorescent proteins

Absorption

Brain

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