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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Fluorescence lifetime imaging through scattering media.

Sebastian Nilsson1, Elias Kristensson2, Marcus Aldén2

  • 1Combustion Physics, Department of Physics, Faculty of Engineering, Lund University, Professorsgatan 1, 223 63, Lund, Sweden. sebastian.nilsson@forbrf.lth.se.

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Structured illumination enhances fluorescence lifetime imaging by improving contrast and removing scattered light. This technique enables accurate molecular identification and concentration estimation even in challenging scattering media.

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Area of Science:

  • Optics and Photonics
  • Biophysical Techniques

Background:

  • Fluorescence lifetime determination is vital for molecular identification, concentration estimation, and temperature sensing.
  • Mixed decay rates and low contrast due to light scattering complicate accurate lifetime measurements.

Purpose of the Study:

  • To develop a method for improving contrast in fluorescence lifetime wide-field imaging.
  • To enable accurate fluorescence lifetime imaging in the presence of scattering media.

Main Methods:

  • Utilized structured illumination to enhance image contrast in fluorescence lifetime wide-field imaging.
  • Employed Dual Imaging Modeling Evaluation (DIME) for lifetime imaging determination.
  • Applied spatial lock-in analysis to remove spurious scattered signals.

Main Results:

  • Successfully enhanced image contrast using structured illumination.
  • Demonstrated the removal of spurious scattered signals via spatial lock-in analysis.
  • Enabled reliable fluorescence lifetime imaging through scattering media.

Conclusions:

  • Structured illumination combined with spatial lock-in analysis overcomes limitations of low contrast and scattering.
  • This approach significantly improves the accuracy and applicability of fluorescence lifetime imaging.