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Proceedings Paper

Hybrid reflecting objectives for deep-tissue functional two-photon imaging
Author(s): Dejan Vučinić; Thomas M. Bartol; Terrence J. Sejnowski
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Paper Abstract

Most microscope objectives commonly used for two-photon imaging of neurons are optimized for high image quality under wide-field illumination and for long working distance, constraints that are at odds with the need for high fluorescence collection efficiency and large field of view dictated by the low fluorescence intensity and the scattering properties of brain tissue. We present a de novo design of an objective intended specifically for deep-tissue functional two-photon imaging. Our design has separate imaging and non-imaging pathways for incident and emitted light, making use of the optical sectioning intrinsic in non-linear fluorescence excitation, which relaxes a number of design constraints. We show through modeling that a twofold to fourfold improvement in fluorescence collection efficiency over traditional objective designs is easily achievable while maintaining nearly diffraction-limited performance within a 200-micron field of view and a long working distance.

Paper Details

Date Published: 31 August 2006
PDF: 7 pages
Proc. SPIE 6288, Current Developments in Lens Design and Optical Engineering VII, 62880U (31 August 2006); doi: 10.1117/12.680952
Show Author Affiliations
Dejan Vučinić, The Salk Institute for Biological Studies (United States)
Thomas M. Bartol, The Salk Institute for Biological Studies (United States)
Terrence J. Sejnowski, The Salk Institute for Biological Studies (United States)
Univ. of California, San Diego (United States)


Published in SPIE Proceedings Vol. 6288:
Current Developments in Lens Design and Optical Engineering VII
Pantazis Z. Mouroulis; Warren J. Smith; R. Barry Johnson, Editor(s)

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