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Updated: Jun 1, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Phasor-based single-molecule fluorescence lifetime imaging using a wide-field photon-counting detector.

R Colyer¹, O Siegmund, A Tremsin

¹Department of Chemistry & Biochemistry, UCLA, Los Angeles, CA.

Proceedings of Spie--The International Society for Optical Engineering

|June 1, 2011

Summary

This study introduces a new wide-field fluorescence lifetime imaging (FLIM) system combined with phasor analysis for faster and more detailed molecular environment studies. The approach enhances biological and material science research by improving data acquisition and analysis.

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Area of Science:

Biophysics
Molecular Imaging
Spectroscopy

Background:

Fluorescence lifetime imaging (FLIM) provides insights into molecular environments and processes like FRET.
Traditional FLIM analysis can be limited by signal requirements and slow acquisition rates.
The phasor approach offers an alternative for analyzing fluorescence lifetime data.

Purpose of the Study:

To present a novel FLIM acquisition hardware (H33D detector) for high temporal and spatial resolution.
To demonstrate the advantages of combining this new hardware with phasor analysis for FLIM data.
To illustrate the application of this combined approach using experimental data from live cells and quantum dots.

Main Methods:

Utilizing a high temporal and spatial resolution wide-field single-photon counting device (H33D detector).
Implementing the phasor approach for fluorescence lifetime analysis.
Acquiring and analyzing FLIM data from live cells and quantum dots.

Main Results:

The H33D detector enables high-resolution FLIM acquisition.
Phasor analysis effectively processes the acquired FLIM data.
The combined approach allows for efficient study of molecular environments in complex biological samples.

Conclusions:

The integration of the H33D detector with phasor analysis represents a significant advancement in FLIM.
This method offers a powerful tool for studying molecular dynamics and interactions.
The approach has broad applicability in biological imaging and materials science.