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Related Experiment Videos

Fluorescence lifetime imaging by asynchronous pump-probe microscopy

C Y Dong1, P T So, T French

  • 1Department of Physics, University of Illinois at Urbana-Champaign 61801, USA. chen@lfd.physics.uiuc.edu

Biophysical Journal
|December 1, 1995
PubMed
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We developed a scanning lifetime fluorescence microscope using a pump-probe technique. This method achieves axial sectioning and enhanced resolution without fast detectors, enabling lifetime-resolved imaging.

Area of Science:

  • Optics and Photonics
  • Biomedical Imaging
  • Fluorescence Microscopy

Background:

  • Conventional fluorescence microscopy faces limitations in resolution and depth penetration.
  • Confocal and two-photon microscopy offer sectioning but can be complex.
  • Lifetime-resolved imaging typically requires high-speed photodetectors.

Purpose of the Study:

  • To develop a novel scanning lifetime fluorescence microscope.
  • To leverage the asynchronous pump-probe technique for enhanced imaging capabilities.
  • To enable lifetime-resolved imaging without high-speed detectors.

Main Methods:

  • Utilized an asynchronous, pump-probe (stimulated emission) approach for microscopy.
  • Generated a cross-correlation signal by overlapping pump and probe lasers.

Related Experiment Videos

  • Measured fluorescence lifetime at high frequencies (up to 6.7 GHz).

    • Main Results:

    • Achieved axial sectioning and improved spatial resolution compared to one-photon microscopy.
    • Demonstrated lifetime-resolved imaging using a low-frequency cross-correlation signal.
    • Successfully imaged fluorescent latex spheres, erythrocytes, and stained fibroblast cells.

    Conclusions:

    • The developed pump-probe microscope offers advantages in resolution and sectioning.
    • This technique provides lifetime-resolved imaging capabilities without specialized fast detectors.
    • The method is applicable to biological samples and advanced fluorescence imaging.