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 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.
Gratton
Enrico
E
University of Illinois at Urbana-Champaign, Laboratory for Fluorescence Dynamics, 1110 West Green Street Urbana, Illinois 61801, USA. Enrico@scs.uiuc.edu
Breusegem
Sophie
S
Sutin
Jason
J
Ruan
Qiaoqiao
Q
Barry
Nicholas
N
D000263:Adenylate Kinase / Q000378:metabolism*; D000465:Algorithms; D000818:Animals; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003198:Computer Simulation; D019793:Fluorescein; D005583:Fourier Analysis; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D039641:Integrin beta Chains / Q000378:metabolism*; D007822:Laryngeal Neoplasms / Q000201:enzymology* / Q000473:pathology; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D029742:Caenorhabditis elegans Proteins; D039641:Integrin beta Chains; D011993:Recombinant Fusion Proteins; D000263:Adenylate Kinase; D019793:Fluorescein