title: Automated cell-specific laser detection and ablation of neural circuits in neonatal brain tissue.
abstract: A key feature of neurodegenerative disease is the pathological loss of neurons that participate in generating behaviour. To investigate network properties of neural circuits and provide a complementary tool to study neurodegeneration in vitro or in situ, we developed an automated cell-specific laser detection and ablation system. The instrument consists of a two-photon and visible-wavelength confocal imaging setup, controlled by executive software, that identifies neurons in preparations based on genetically encoded fluorescent proteins or Ca(2+) imaging, and then sequentially ablates cell targets while monitoring network function concurrently. Pathological changes in network function can be directly attributed to ablated cells, which are logged in real time. Here, we investigated brainstem respiratory circuits to demonstrate single-cell precision in ablation during physiological network activity, but the technique could be applied to interrogate network properties in neural systems that retain network functionality in reduced preparations in vitro or in situ.
authors: Department of Applied Science, McGlothlin-Street Hall, Room 318, The College of William & Mary, Williamsburg, VA 23187-8795, USA.
authors: Xueying
authors: X
authors: Wang
authors: John A
authors: JA
authors: Hayes
authors: Maria Cristina D
authors: MC
authors: Picardo
authors: Christopher A
authors: CA
authors: Del Negro
mesh_terms: D000465:Algorithms; D000814:Aniline Compounds; D000818:Animals; D000831:Animals, Newborn; D001933:Brain Stem / Q000502:physiology*; D002118:Calcium / Q000502:physiology; D005260:Female; D005456:Fluorescent Dyes; D018398:Homeodomain Proteins / Q000502:physiology; D053685:Laser Therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D009474:Neurons / Q000502:physiology*; D011247:Pregnancy; D012984:Software; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; C103403:Dbx1 protein, mouse; D005456:Fluorescent Dyes; D018398:Homeodomain Proteins; D014966:Xanthenes; C561295:fluo-8 AM; D002118:Calcium