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Molecular probes for fluorescence lifetime imaging.

Pinaki Sarder1, Dolonchampa Maji1, Samuel Achilefu1

  • 1†Departments of Radiology, ‡Biomedical Engineering, and §Biochemistry and Molecular Biophysics, Washington University School of Medicine, 4525 Scott Avenue, St. Louis, Missouri 63110, United States.

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Fluorescence lifetime (FLT) imaging offers a powerful alternative to intensity-based methods for visualizing biological processes. This technique overcomes limitations and provides deeper insights into cellular and tissue environments.

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Area of Science:

  • Biophotonics
  • Molecular Imaging
  • Cellular Biology

Background:

  • Fluorescence intensity imaging is standard for visualizing biological processes but has limitations like concentration dependency.
  • Fluorescence lifetime (FLT) imaging provides complementary information, overcoming intensity-based measurement challenges.
  • FLT imaging enables evaluation of molecular interactions and biochemical status in cellular microenvironments.

Purpose of the Study:

  • To review recent advances in FLT imaging of molecular probes for biological applications.
  • To highlight the potential of FLT imaging in cellular and small animal models.
  • To identify challenges and future directions for FLT molecular probe development.

Main Methods:

  • Utilizing a wide array of dyes with stable or environment-responsive FLTs for information multiplexing.
  • Applying FLT spectroscopy and microscopy for biological studies.
  • Developing FLT imaging techniques for in vivo applications in small animal models.

Main Results:

  • FLT imaging allows for reliable assessment of molecular interactions and microenvironment status.
  • Information multiplexing is achievable without ratiometric spectral imaging.
  • FLT imaging is increasingly applied to cells and small animal models of human diseases.

Conclusions:

  • FLT imaging is a valuable tool for understanding biological processes and disease states.
  • Further development of FLT imaging techniques is needed for widespread in vivo applications.
  • FLT molecular probes hold significant potential for addressing diverse biological questions.