title: Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices.
abstract: We developed a two-photon microscope optimized for physiologically manipulating single neurons through their postsynaptic receptors. The optical layout fulfills the stringent design criteria required for high-speed, high-resolution imaging in scattering brain tissue with minimal photodamage. We detail the practical compensation of spectral and temporal dispersion inherent in fast laser beam scanning with acousto-optic deflectors, as well as a set of biological protocols for visualizing nearly diffraction-limited structures and delivering physiological synaptic stimuli. The microscope clearly resolves dendritic spines and evokes electrophysiological transients in single neurons that are similar to endogenous responses. This system enables the study of multisynaptic integration and will assist our understanding of single neuron function and dendritic computation.
authors.affiliation: Baylor College of Medicine, Department of Neuroscience, One Baylor Plaza, Houston, Texas 77030, USA.
authors.forename: Bradley E
authors.initials: BE
authors.lastname: Losavio
authors.forename: Vijay
authors.initials: V
authors.lastname: Iyer
authors.forename: Peter
authors.initials: P
authors.lastname: Saggau
mesh_terms: D000818:Animals; D003712:Dendrites / Q000502:physiology; D006624:Hippocampus / Q000502:physiology*; D008297:Male; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D008959:Models, Neurological; D009474:Neurons / Q000502:physiology*; D018377:Neurotransmitter Agents / Q000502:physiology*; D010782:Photolysis; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018377:Neurotransmitter Agents