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Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue

Chaigneau, E. and Wright, A. J. and Poland, S. P. and Girkin, J. M. and Silver, R. A. (2011) Impact of wavefront distortion and scattering on 2-photon microscopy in mammalian brain tissue. Optics Express, 19 (23). pp. 22755-22774. ISSN 1094-4087

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Abstract

Two-photon (2P) microscopy is widely used in neuroscience, but the optical properties of brain tissue are poorly understood. We have investigated the effect of brain tissue on the 2P point spread function (PSF(2P)) by imaging fluorescent beads through living cortical slices. By combining this with measurements of the mean free path of the excitation light, adaptive optics and vector-based modeling that includes phase modulation and scattering, we show that tissue-induced wavefront distortions are the main determinant of enlargement and distortion of the PSF2P at intermediate imaging depths. Furthermore, they generate surrounding lobes that contain more than half of the 2P excitation. These effects reduce the resolution of fine structures and contrast and they, together with scattering, limit 2P excitation. Our results disentangle the contributions of scattering and wavefront distortion in shaping the cortical PSF2P, thereby providing a basis for improved 2P microscopy.

Item type: Article
ID code: 41713
Keywords: systems, adaptive optics, implementation, samples, specimen-induced aberrations, electromagnetic diffraction, fluorescence microscopy, multiphoton microscopy, image field, wavefront distortion , scattering, 2-photon microscopy , mammalian brain tissue , Pharmacy and materia medica, Neuroscience. Biological psychiatry. Neuropsychiatry, Atomic and Molecular Physics, and Optics
Subjects: Medicine > Pharmacy and materia medica
Medicine > Internal medicine > Neuroscience. Biological psychiatry. Neuropsychiatry
Department: Faculty of Science > Institute of Photonics
Related URLs:
Depositing user: Pure Administrator
Date Deposited: 24 Oct 2012 16:59
Last modified: 27 Mar 2014 10:36
URI: http://strathprints.strath.ac.uk/id/eprint/41713

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