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Photoluminescence: Fluorescence and Phosphorescence01:23

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Related Experiment Video

Updated: Nov 4, 2025

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
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Two-Photon Phosphorescence Lifetime Microscopy.

Nastaran Abbasizadeh1,2, Joel A Spencer3,4

  • 1Department of Bioengineering, University of California Merced, Merced, CA, USA.

Advances in Experimental Medicine and Biology
|May 30, 2021
PubMed
Summary
This summary is machine-generated.

Two-photon Phosphorescence Lifetime Microscopy (2PLM) offers functional imaging by measuring oxygen levels in biological samples. This technique uses phosphorescence lifetime to monitor oxygen tension, aiding in understanding human health and disease.

Keywords:
2PLMOxygen sensingPQMPhosphorescence lifetime microscopyPhosphorescence quenchingTissue oxygenationTwo-photon excitation

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Area of Science:

  • Biomedical Optics
  • Molecular Imaging
  • Physiology

Background:

  • Two-photon Phosphorescence Lifetime Microscopy (2PLM) is an advanced nonlinear optical imaging technique.
  • It differs from fluorescence imaging by utilizing the longer-lived triplet excited state for phosphorescence emission (microseconds to milliseconds).
  • This long lifetime makes 2PLM sensitive to oxygen quenching, enabling oxygen tension measurements.

Purpose of the Study:

  • To provide background on 2PLM development and principles.
  • To discuss oxygen sensing methods and concepts using 2PLM.
  • To review biological applications and impact on human health research.

Main Methods:

  • Utilizes two-photon excitation with near-infrared wavelengths for deep tissue penetration and low phototoxicity.
  • Measures phosphorescence lifetime, which is quenched by local oxygen concentration.
  • Generates 3D optical sections within thick biological specimens.

Main Results:

  • 2PLM provides both probe distribution (intensity) and local oxygen tension (lifetime quenching) information.
  • The technique allows for longitudinal imaging and monitoring of oxygen in living organisms.
  • Demonstrates sensitivity to oxygen levels crucial for biological processes.

Conclusions:

  • 2PLM is a powerful functional imaging tool for biological research.
  • It enhances understanding of oxygen's role in human health and disease.
  • Further improvements in 2PLM characteristics are key for broader applications.