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Fluorescence lifetime imaging.

J R Lakowicz1, H Szmacinski, K Nowaczyk

  • 1Department of Biological Chemistry, University of Maryland School of Medicine, Baltimore 21201.

Analytical Biochemistry
|May 1, 1992
PubMed
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We developed a new fluorescence imaging method using fluorescence lifetime, not intensity, for contrast. This technique enables rapid, simultaneous pixel data acquisition for advanced bioscience applications.

Area of Science:

  • Biophysics
  • Optical Imaging
  • Spectroscopy

Background:

  • Traditional fluorescence imaging relies on fluorophore concentration or intensity.
  • This limits the ability to probe specific molecular environments and processes.
  • A method to extract contrast from fluorescence decay dynamics is needed.

Purpose of the Study:

  • To introduce a novel fluorescence imaging methodology based on fluorescence lifetime.
  • To demonstrate its capability for biochemical and physical imaging.
  • To enable faster image acquisition and visualization of unique features.

Main Methods:

  • Utilizing a gain-modulated image intensifier to acquire phase-sensitive images.
  • Employing a slow-scan CCD camera for image collection.

Related Experiment Videos

  • Calculating phase angle and modulation from a series of images with phase shifts to create lifetime images.

    • Main Results:

    • Experimentally verified the method with standard fluorophores (1-10 ns lifetimes).
    • Generated lifetime images of Yt-base quenched by acrylamide, demonstrating environmental sensitivity.
    • Developed a faster imaging procedure for rapid visualization of distinct decay times.

    Conclusions:

    • The developed fluorescence lifetime imaging (FLIM) method provides contrast based on decay time, not intensity.
    • This technique offers simultaneous pixel data acquisition, enhancing imaging speed.
    • FLIM has broad potential applications in biosciences for chemical and physical imaging.