Overcoming the fundamental limitation of frequency-domain fluorescence lifetime imaging microscopy spatial resolution

Xiaotong Yuan; Varun Mannam; Yide Zhang; Scott Howard

doi:10.1117/12.2577284

5 March 2021 Overcoming the fundamental limitation of frequency-domain fluorescence lifetime imaging microscopy spatial resolution

Xiaotong Yuan, Varun Mannam, Yide Zhang, Scott Howard

Author Affiliations +

Proceedings Volume 11650, Single Molecule Spectroscopy and Superresolution Imaging XIV; 116500M (2021) https://doi.org/10.1117/12.2577284
Event: SPIE BiOS, 2021, Online Only

Abstract

We propose and demonstrate the first analytical model of the spatial resolution of frequency-domain (FD) fluorescence lifetime imaging microscopy (FLIM) that explains how it is fundamentally different with the common resolution limit of the conventional fluorescence microscopy. Frequency modulation (FM) capture effect is also observed by the model, which results in distorted FLIM measurements. A super-resolution FLIM approach based on a localization-based technique, super-resolution radial fluctuations (SRRF), is presented. In this approach, we separately process the intensity and lifetime to generate a super-resolution FLIM composite image. The capability of the approach is validated both numerically and experimentally in fixed cells sample.

Conference Presentation

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Xiaotong Yuan, Varun Mannam, Yide Zhang, and Scott Howard "Overcoming the fundamental limitation of frequency-domain fluorescence lifetime imaging microscopy spatial resolution", Proc. SPIE 11650, Single Molecule Spectroscopy and Superresolution Imaging XIV, 116500M (5 March 2021); https://doi.org/10.1117/12.2577284

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