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Fluorescence Lifetime Macro Imager for Biomedical Applications
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Fluorescence Lifetime Macro Imager for Biomedical Applications.

Rajannya Sen1, Alexander V Zhdanov2, Ciaran Devoy3

  • 1School of Biochemistry and Cell Biology, University College Cork; rajannya.sen@ucc.ie.

Journal of Visualized Experiments : Jove
|April 24, 2023
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Summary

A new photoluminescence lifetime imager maps molecular oxygen (O2) concentration in tissues and materials. This fast, sensitive system provides accurate O2 mapping for biological and material science applications.

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Area of Science:

  • Biomedical Optics
  • Molecular Imaging
  • Sensor Technology

Background:

  • Phosphorescence lifetime imaging microscopy (PLIM) is crucial for studying molecular oxygen (O2) levels.
  • Existing PLIM platforms often face limitations in accuracy, flexibility, and ease of use.
  • Accurate O2 mapping is vital for understanding cellular metabolism and disease states.

Purpose of the Study:

  • To develop and validate a novel, high-sensitivity photoluminescence lifetime imager for precise O2 concentration mapping.
  • To demonstrate the system's capability in imaging O2 levels in diverse samples, including live tissues and O2-sensitive materials.
  • To overcome the limitations of current PLIM technologies, enhancing accuracy and usability.

Main Methods:

  • Utilized a Timepix3 camera (Tpx3Cam) integrated with an opto-mechanical adaptor and image intensifier.
  • Employed nanoparticle-based near-infrared probes (NanO2-IR) for O2 sensing, with specific excitation and emission wavelengths.
  • Adapted the system for O2-sensitive Pt-porphyrin dyes using different excitation and emission filters.

Main Results:

  • The developed PLIM imager achieved fast (≤20 s) and accurate mapping of O2 levels in intestinal tissue samples.
  • Demonstrated high-intensity phosphorescence signals and stable lifetime values from various biological and solid-state samples.
  • Successfully imaged hypoxia in grafted tumors in animal models and re-configured for different phosphorescent probes.

Conclusions:

  • The novel PLIM imager offers accurate, quantitative O2 concentration measurements and 2D mapping capabilities.
  • The system provides a versatile and sensitive tool for metabolic imaging of ex vivo and in vivo samples.
  • This technology advances O2 sensing applications in biomedical research and material science.