Research Papers

Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices

[+] Author Affiliations
Bradley E. Losavio

Baylor College of Medicine, Department of Neuroscience, One Baylor Plaza, Houston, Texas 77030

Vijay Iyer

Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147

Peter Saggau

Baylor College of Medicine, Department of Neuroscience and Department of Molecular Physiology and Biophysics, One Baylor Plaza, Houston, Texas 77030

J. Biomed. Opt. 14(6), 064033 (December 31, 2009). doi:10.1117/1.3275468
History: Received April 14, 2009; Revised September 18, 2009; Accepted October 15, 2009; Published December 31, 2009; January 12, 2010
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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.

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© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Bradley E. Losavio ; Vijay Iyer and Peter Saggau
"Two-photon microscope for multisite microphotolysis of caged neurotransmitters in acute brain slices", J. Biomed. Opt. 14(6), 064033 (December 31, 2009). ; http://dx.doi.org/10.1117/1.3275468


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