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Automated cell‐specific laser detection and ablation of neural circuits in neonatal brain tissue

•  We developed an automated system that detects neurons belonging to specific populations in vitro or in situ, maps their physical locations in three‐dimensional tissue specimens and then laser ablates the cell ‘targets’ one at a time, in sequence, while monitoring neural population activity electrophysiologically. •  Two‐photon Ca2+ imaging and image processing routines detect and validate target neurons based on rhythmic Ca2+ fluorescence activity patterns. •  Visible‐wavelength confocal imaging and image processing routines detect and validate target neurons that express genetically encoded fluorescent proteins. •  High‐intensity two‐photon spot scanning vaporizes target neurons with specificity while minimizing damage to neighbouring tissue. •  Physiological monitoring of network function is performed before, during and after the cell‐specific laser ablations to measure the effects on network functionality in real time.