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Updated: Dec 27, 2025

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
High temporal resolution and polarization resolved fluorescence lifetime measurements through stimulated emission
Khalil Ur Rehman1, Subir Das1, Yih-Fan Chen1
1Institute of Biophotonics, National Yang-Ming University, Taipei, 11221, Taiwan.
Abstract:
We have implemented polarization-resolved fluorescence lifetime measurement through stimulated emission based pump-probe technique, which promises much higher temporal resolution (∼4 ps) than conventional time-correlated single-photon counting (TCSPC). The depolarization of ATTO 647N fluorescent dye is resolved through anisotropy fluorescence lifetime measurements, with variable time delay introduced between the pump and the probe beams. Importantly, the polarization anisotropy measurement and the corresponding rotational correlation time characterization of the fluorescent dye are carried out at various temperatures. We have also demonstrated the need of high temporal resolution via hetero Förster energy transfer (Hetero-FRET) through the interaction between the gold nanorods (GNRs) and the fluorescent dye ATTO 647N. Notably, our results compare highly favorably with conventional TCSPC method, which is rather limited in temporal resolution, for the above characterization. Additionally, this technique is applicable even under ambient light while being very cost-effective and robust.
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