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Multispectral fluorescence lifetime imaging by TCSPC.

Wolfgang Becker1, Axel Bergmann, Christoph Biskup

  • 1Becker & Hickl, Nahmitzer Damm 30, 12277 Berlin, Germany. becker@becker-hickl.de

Microscopy Research and Technique
|March 30, 2007
PubMed
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We developed a new fluorescence lifetime imaging technique for multiphoton microscopes. This method offers simultaneous spectral and temporal resolution, improving autofluorescence imaging of tissue with high efficiency.

Area of Science:

  • Biophotonics
  • Microscopy
  • Spectroscopy

Background:

  • Multiphoton microscopy is crucial for deep tissue imaging.
  • Simultaneous spectral and temporal resolution in fluorescence imaging is challenging.
  • Autofluorescence imaging provides valuable insights into tissue physiology and pathology.

Purpose of the Study:

  • To present a novel fluorescence lifetime imaging technique.
  • To achieve simultaneous spectral and temporal resolution in multiphoton microscopy.
  • To demonstrate the technique's utility for autofluorescence imaging of tissue.

Main Methods:

  • Utilized a fiber bundle to relay the microscope's back-aperture image.
  • Employed a spectrograph and a 16-anode photomultiplier tube (PMT) module.

Related Experiment Videos

  • Implemented multidimensional time-correlated single photon counting (TCSPC) for data acquisition.

    • Main Results:

    • Achieved near-ideal photon counting efficiency.
    • Demonstrated the capability to resolve double-exponential decay functions.
    • Successfully applied the technique for autofluorescence imaging of tissue.

    Conclusions:

    • The developed technique is compatible with standard multiphoton microscopes and direct detection.
    • The method provides high-resolution spectral and temporal information for fluorescence imaging.
    • This technique enhances the diagnostic potential of autofluorescence imaging in biological tissues.