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

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 approache...

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Bibliographic Details
Published inJournal of biomedical optics Vol. 8; no. 3; p. 381
Main Authors Gratton, Enrico, Breusegem, Sophie, Sutin, Jason, Ruan, Qiaoqiao, Barry, Nicholas
Format Journal Article
LanguageEnglish
Published United States 01.07.2003
Subjects
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
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Summary: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.
ISSN:1083-3668
1560-2281
DOI:10.1117/1.1586704