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Fluorescence Lifetime Macro Imager for Biomedical Applications
06:01

Fluorescence Lifetime Macro Imager for Biomedical Applications

Published on: April 7, 2023

Fluorescence lifetime optical projection tomography.

James McGinty1, Khadija B Tahir, Romain Laine

  • 1Department of Physics, Imperial College London, Prince Consort Road, South Kensington, London, SW7 2BW, UK. james.mcginty@imperial.ac.uk

Journal of Biophotonics
|April 4, 2009
PubMed
Summary
This summary is machine-generated.

We developed a quantitative 3-D fluorescence lifetime tomography technique for cleared specimens. This method accurately maps fluorescence lifetime distributions, distinguishing labels from autofluorescence in biological samples.

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Area of Science:

  • Biomedical Imaging
  • Optical Physics
  • Molecular Biology

Background:

  • Quantitative fluorescence imaging is crucial for biological research.
  • Existing methods struggle with intensity-based artifacts and autofluorescence.
  • Accurate 3-D mapping of fluorescence lifetime is needed for complex samples.

Purpose of the Study:

  • To develop and validate a quantitative fluorescence projection tomography (FPT) technique.
  • To measure 3-D fluorescence lifetime distribution in optically cleared biological specimens.
  • To overcome limitations of intensity-based fluorescence imaging.

Main Methods:

  • Acquisition of time-gated wide-field images at various projection angles and time delays.
  • Reconstruction of 3-D time-gated intensity distributions using filtered back projection.
  • Determination of fluorescence lifetime via iterative mono-exponential decay fitting on reconstructed planes.

Main Results:

  • Demonstration of a quantitative 3-D fluorescence lifetime tomography technique.
  • Successful reconstruction of fluorescence lifetime in optically cleared specimens up to 1 cm.
  • Robustness against intensity artifacts due to the ratiometric nature of fluorescence lifetime.

Conclusions:

  • The developed FPT technique provides quantitative 3-D fluorescence lifetime mapping.
  • This method effectively differentiates extrinsic labels from autofluorescence in biological samples.
  • The technique shows promise for advanced imaging of labeled structures in complex biological systems, such as mouse embryos.