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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Fourier Multiplexed Fluorescence Lifetime Imaging.

Leilei Peng1

  • 1Wyant College of Optical Science, University of Arizona, Tucson, AZ, USA. lpeng@optics.arizona.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2021
PubMed
Summary
This summary is machine-generated.

Fourier multiplexed FLIM (FmFLIM) enables parallel detection of multiple fluorescence labels. This advanced imaging technique allows for multiplexed 3D lifetime imaging in biological samples like cells, tissues, and embryos.

Keywords:
Förster resonance energy transferLifetime imagingMultiplexed imaging

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Area of Science:

  • Biophotonics and advanced microscopy techniques.

Background:

  • Fluorescence lifetime imaging microscopy (FLIM) is a key functional imaging modality in biosciences.
  • Existing FLIM methods may face limitations in parallel detection of multiple fluorescence labels.

Purpose of the Study:

  • To introduce and detail Fourier multiplexed FLIM (FmFLIM) as a method for parallel lifetime detection.
  • To demonstrate the application of FmFLIM in advanced 3D imaging of biological specimens.

Main Methods:

  • Utilizes frequency-domain lifetime measurements based on Fourier (frequency) multiplexing principles.
  • Integrates FmFLIM with confocal scanning microscopy for multiplexed 3D imaging of cells and tissues.
  • Combines FmFLIM with scanning laser tomography for 3D multiplex lifetime imaging of whole embryos and thick tissues.

Main Results:

  • Achieves parallel lifetime detection for multiple fluorescence labels.
  • Enables multiplexed 3D lifetime imaging of cellular and tissue structures.
  • Facilitates deep tissue imaging and whole embryo analysis using integrated tomography.

Conclusions:

  • FmFLIM offers a powerful approach for simultaneous multi-label lifetime measurements.
  • The integration of FmFLIM with scanning microscopy and tomography expands its utility for complex biological imaging.
  • This technique enhances the capability for high-content functional imaging in biosciences.