Super-resolution multiphoton frequency-domain fluorescence lifetime imaging microscopy by generalized stepwise optical saturation (GSOS)

Yide Zhang; David Benirschke; Ola Abdalsalam; Scott S. Howard

doi:10.1117/12.2507663

22 February 2019 Super-resolution multiphoton frequency-domain fluorescence lifetime imaging microscopy by generalized stepwise optical saturation (GSOS)

Yide Zhang, David Benirschke, Ola Abdalsalam, Scott S. Howard

Author Affiliations +

Proceedings Volume 10884, Single Molecule Spectroscopy and Superresolution Imaging XII; 108840C (2019) https://doi.org/10.1117/12.2507663
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

We present the first experimental demonstration of super-resolution multiphoton frequency-domain (FD) fluorescence lifetime imaging microscopy (FLIM). This is obtained through a novel microscopy technique called generalized stepwise optical saturation (GSOS). GSOS√utilizes the linear combination of M steps of raw images to improve the imaging resolution by a factor of √M . Here, a super-resolution multiphoton FD-FLIM is demonstrated on various samples, including fixed cells and biological tissues, with a custom-built two-photon FD-FLIM microscope. We demonstrate simultaneous super-resolution intensity and fluorescence lifetime images of a variety of cell cultures and ex vivo tissues. Combined with multiphoton excitation, the proposed GSOS microscopy is able to generate super-resolution FLIM images deep in scattering samples.

Conference Presentation

Citation Download Citation

Yide Zhang, David Benirschke, Ola Abdalsalam, and Scott S. Howard "Super-resolution multiphoton frequency-domain fluorescence lifetime imaging microscopy by generalized stepwise optical saturation (GSOS)", Proc. SPIE 10884, Single Molecule Spectroscopy and Superresolution Imaging XII, 108840C (22 February 2019); https://doi.org/10.1117/12.2507663

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