title: Laser-Mediated Microlesions in Mouse Neocortex to Investigate Neuronal Degeneration and Regeneration.
abstract: In vivo two-photon (2P) imaging enables neural circuitry to be repeatedly visualized in both normal conditions and following trauma. This protocol describes how laser-mediated neuronal microlesions can be created in the cerebral cortex using an ultrafast laser without causing a significant inflammatory reaction or compromising the blood-brain barrier. Furthermore, directives are provided for the acute and chronic in vivo imaging of the lesion site, as well as for post-hoc analysis of the lesion site in fixed tissue, which can be correlated with the live imaging phase.
authors.affiliation: Lilly UK, Windlesham, United Kingdom.|These authors contributed equally to this work.
authors.forename: Johanna
authors.initials: J
authors.lastname: Jackson
authors.affiliation: Wicking Dementia Research and Education Centre, University of Tasmania, Hobart, Australia.|These authors contributed equally to this work.
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Canty
authors.affiliation: Australian Regenerative Medicine Institute, Monash University, Clayton, Australia.
authors.forename: Lieven
authors.initials: L
authors.lastname: Huang
authors.affiliation: MRC Clinical Sciences Centre, Imperial College London, London, United Kingdom.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Paola
mesh_terms: D000818:Animals; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D007834:Lasers / Q000009:adverse effects*; D051379:Mice; D019579:Neocortex / Q000166:cytology* / Q000293:injuries*; D009410:Nerve Degeneration / Q000209:etiology*; D009416:Nerve Regeneration / Q000502:physiology*; D059906:Neuroimaging; D009474:Neurons / Q000378:metabolism / Q000502:physiology*; D009924:Organ Culture Techniques
chemical_list: D049452:Green Fluorescent Proteins
keywords: 2-photon microscopy; axons; axotomy; cortex; glial scar; laser ablation; live imaging; microlesion; synapses