Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods

J Biomed Opt. 2003 Jul;8(3):381-90. doi: 10.1117/1.1586704.

Abstract

Fluorescence lifetime images are obtained with the laser scanning microscope using two methods: the time-correlated single-photon counting method and the frequency-domain method. In the same microscope system, we implement both methods. We perform a comparison of the performance of the two approaches in terms of signal-to-noise ratio (SNR) and the speed of data acquisition. While in our practical implementation the time-correlated single-photon counting technique provides a better SNR for low-intensity images, the frequency-domain method is faster and provides less distortion for bright samples.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Adenylate Kinase / metabolism*
  • Algorithms*
  • Animals
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / metabolism
  • Cell Line, Tumor
  • Cells, Cultured
  • Computer Simulation
  • Fluorescein
  • Fourier Analysis
  • Humans
  • Image Enhancement / methods*
  • Integrin beta Chains / metabolism*
  • Laryngeal Neoplasms / enzymology*
  • Laryngeal Neoplasms / pathology
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Caenorhabditis elegans Proteins
  • Integrin beta Chains
  • Recombinant Fusion Proteins
  • Adenylate Kinase
  • Fluorescein