Dr. Dejan Vucinic Profile

Dr. Dejan Vucinic

at Stanford Univ

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Proceedings Article | 31 August 2006 Paper

Hybrid reflecting objectives for deep-tissue functional two-photon imaging

Dejan Vučinić, Thomas Bartol, Terrence Sejnowski

Proceedings Volume 6288, 62880U (2006) https://doi.org/10.1117/12.680952

KEYWORDS: Objectives, Luminescence, Photons, Mirrors, Two photon imaging, Microscopes, Tissues, Tissue optics, Scattering, Image quality

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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.

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