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Related Experiment Video

Updated: Mar 26, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Multi-target spectrally resolved fluorescence lifetime imaging microscopy.

Thomas Niehörster¹, Anna Löschberger¹, Ingo Gregor²

¹Department of Biotechnology &Biophysics, Julius Maximilian University of Würzburg, Würzburg, Germany.

|January 26, 2016

Summary

This summary is machine-generated.

This study presents a new method for identifying multiple fluorescent molecules in cells using spectrally resolved fluorescence lifetime imaging microscopy (sFLIM). This technique enables simultaneous visualization and imaging of up to nine target molecules with high precision.

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Last Updated: Mar 26, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

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Published on: December 29, 2017

Area of Science:

Biophysics
Microscopy
Molecular Imaging

Background:

Multidimensional fluorescence signals present challenges for identifying individual fluorophores.
Existing methods struggle with simultaneous detection of multiple targets in complex biological samples.

Purpose of the Study:

To develop an efficient pattern-matching technique for identifying fluorophore ratios in complex fluorescence signals.
To enable simultaneous multi-target imaging using spectrally resolved fluorescence lifetime imaging microscopy (sFLIM).

Main Methods:

Utilized alternating pulsed laser excitation at three wavelengths and time-resolved detection across 32 channels.
Employed reference fluorescence decay and spectral signature patterns for probe identification.
Applied spectrally resolved fluorescence lifetime imaging microscopy (sFLIM) for data acquisition.

Main Results:

Successfully visualized up to nine different target molecules simultaneously in mouse C2C12 cells.
Achieved multi-target imaging of three different molecules using the same fluorophore in human U2OS cells by exploiting environmental sensitivities.
Demonstrated sFLIM's capability for super-resolution multi-target imaging when combined with stimulated emission depletion (STED).

Conclusions:

The developed pattern-matching technique enhances the efficiency of fluorophore identification in complex biological systems.
sFLIM is a powerful tool for high-content, multi-target molecular imaging in cellular environments.
sFLIM combined with STED offers potential for advanced super-resolution imaging applications.