Three-dimensional deep tissue multiphoton frequency-domain fluorescence lifetime imaging microscopy via phase multiplexing and adaptive optics

Yide Zhang; Ian H. Guldner; Evan L. Nichols; David Benirschke; Cody J. Smith; Siyuan Zhang; Scott S. Howard

doi:10.1117/12.2510674

22 February 2019 Three-dimensional deep tissue multiphoton frequency-domain fluorescence lifetime imaging microscopy via phase multiplexing and adaptive optics

Yide Zhang, Ian H. Guldner, Evan L. Nichols, David Benirschke, Cody J. Smith, Siyuan Zhang, Scott S. Howard

Author Affiliations +

Proceedings Volume 10882, Multiphoton Microscopy in the Biomedical Sciences XIX; 108822H (2019) https://doi.org/10.1117/12.2510674
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

We propose and demonstrate a novel multiphoton frequency-domain fluorescence lifetime imaging microscopy (MPM-FD-FLIM) system that is able to generate 3D lifetime images in deep scattering tissues. The imaging speed of FD-FLIM is improved using phase multiplexing, where the fluorescence signal is split and mixed with the reference signal from the laser in a multiplexing manner. The system allows for easy generation of phasor plots, which not only address multi-exponential decay problems but also clearly represent the dynamics of the fluorophores being investigated. Lastly, a sensorless adaptive optics setup is used for FLIM imaging in deep scattering tissues. The capability of the system is demonstrated in fixed and living animal models, including mice and zebrafish.

Citation Download Citation

Yide Zhang, Ian H. Guldner, Evan L. Nichols, David Benirschke, Cody J. Smith, Siyuan Zhang, and Scott S. Howard "Three-dimensional deep tissue multiphoton frequency-domain fluorescence lifetime imaging microscopy via phase multiplexing and adaptive optics", Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 108822H (22 February 2019); https://doi.org/10.1117/12.2510674

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