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

Dejan Vučinić; Thomas M. Bartol Jr.; Terrence J. Sejnowski

doi:10.1117/12.680952

31 August 2006 Hybrid reflecting objectives for deep-tissue functional two-photon imaging

Dejan Vučinić, Thomas M. Bartol Jr., Terrence J. Sejnowski

Proceedings Volume 6288, Current Developments in Lens Design and Optical Engineering VII; 62880U (2006) https://doi.org/10.1117/12.680952
Event: SPIE Optics + Photonics, 2006, San Diego, California, United States

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.

Citation Download Citation

Dejan Vučinić, Thomas M. Bartol Jr., and Terrence J. Sejnowski "Hybrid reflecting objectives for deep-tissue functional two-photon imaging", Proc. SPIE 6288, Current Developments in Lens Design and Optical Engineering VII, 62880U (31 August 2006); https://doi.org/10.1117/12.680952

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KEYWORDS

Objectives

Luminescence

Mirrors

Photons

Two photon imaging

Microscopes

Tissues

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