| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Biophys J, April 2000, p. 2127-2137, Vol. 78, No. 4
Cell Biophysics Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, England
Global analysis techniques are described for frequency
domain fluorescence lifetime imaging microscopy (FLIM) data. These algorithms exploit the prior knowledge that only a limited number of
fluorescent molecule species whose lifetimes do not vary spatially are
present in the sample. Two approaches to implementing the lifetime
invariance constraint are described. In the lifetime invariant fit
method, each image in the lifetime image sequence is spatially averaged
to obtain an improved signal-to-noise ratio. The lifetime estimations
from these averaged data are used to recover the fractional
contribution to the steady-state fluorescence on a pixel-by-pixel basis
for each species. The second, superior, approach uses a global analysis
technique that simultaneously fits the fractional contributions in all
pixels and the spatially invariant lifetimes. In frequency domain FLIM
the maximum number of lifetimes that can be fit with the global
analysis method is twice the number of lifetimes that can be fit with
conventional approaches. As a result, it is possible to discern two
lifetimes with a single-frequency FLIM setup. The algorithms were
tested on simulated data and then applied to separate the cellular
distributions of coexpressed green fluorescent proteins in living cells.
Biophys J, April 2000, p. 2127-2137, Vol. 78, No. 4
© 2000 by the Biophysical Society 0006-3495/00/04/2127/11 $2.00
This article has been cited by other articles:
 |
|
 |
|
  S. Padilla-Parra, N. Auduge, M. Coppey-Moisan, and M. Tramier Quantitative FRET Analysis by Fast Acquisition Time Domain FLIM at High Spatial Resolution in Living Cells Biophys. J., September 15, 2008; 95(6): 2976 - 2988. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G.-J. Kremers, E. B. van Munster, J. Goedhart, and T. W. J. Gadella Jr. Quantitative Lifetime Unmixing of Multiexponentially Decaying Fluorophores Using Single-Frequency Fluorescence Lifetime Imaging Microscopy Biophys. J., July 1, 2008; 95(1): 378 - 389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Digman, V. R. Caiolfa, M. Zamai, and E. Gratton The Phasor Approach to Fluorescence Lifetime Imaging Analysis Biophys. J., January 15, 2008; 94(2): L14 - L16. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Neves-Petersen, S. Klitgaard, A. S. L. Carvalho, S. B. Petersen, M. R. Aires de Barros, and E. P. e Melo Photophysics and Photochemistry of Horseradish Peroxidase A2 upon Ultraviolet Illumination Biophys. J., March 15, 2007; 92(6): 2016 - 2027. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Esposito, H. C. Gerritsen, and F. S. Wouters Fluorescence Lifetime Heterogeneity Resolution in the Frequency Domain by Lifetime Moments Analysis Biophys. J., December 1, 2005; 89(6): 4286 - 4299. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Keese, R. J. Magdeburg, T. Herzog, T. Hasenberg, M. Offterdinger, R. Pepperkok, J. W. Sturm, and P. I. H. Bastiaens Imaging Epidermal Growth Factor Receptor Phosphorylation in Human Colorectal Cancer Cells and Human Tissues J. Biol. Chem., July 29, 2005; 280(30): 27826 - 27831. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. K. Stepinac, S. R. Chamot, E. Rungger-Brandle, P. Ferrez, J.-L. Munoz, H. van den Bergh, C. E. Riva, C. J. Pournaras, and G. A. Wagnieres Light-Induced Retinal Vascular Damage by Pd-porphyrin Luminescent Oxygen Probes Invest. Ophthalmol. Vis. Sci., March 1, 2005; 46(3): 956 - 966. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Parsons, J. Monypenny, S. M. Ameer-Beg, T. H. Millard, L. M. Machesky, M. Peter, M. D. Keppler, G. Schiavo, R. Watson, J. Chernoff, et al. Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells Mol. Cell. Biol., March 1, 2005; 25(5): 1680 - 1695. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Peter, S. M. Ameer-Beg, M. K. Y. Hughes, M. D. Keppler, S. Prag, M. Marsh, B. Vojnovic, and T. Ng Multiphoton-FLIM Quantification of the EGFP-mRFP1 FRET Pair for Localization of Membrane Receptor-Kinase Interactions Biophys. J., February 1, 2005; 88(2): 1224 - 1237. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Pelet, M. J. R. Previte, L. H. Laiho, and P. T. C. So A Fast Global Fitting Algorithm for Fluorescence Lifetime Imaging Microscopy Based on Image Segmentation Biophys. J., October 1, 2004; 87(4): 2807 - 2817. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. H. A. Clayton, Q. S. Hanley, D. J. Arndt-Jovin, V. Subramaniam, and T. M. Jovin Dynamic Fluorescence Anisotropy Imaging Microscopy in the Frequency Domain (rFLIM) Biophys. J., September 1, 2002; 83(3): 1631 - 1649. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. J. Verveer, F. S. Wouters, A. R. Reynolds, and P. I. H. Bastiaens Quantitative Imaging of Lateral ErbB1 Receptor Signal Propagation in the Plasma Membrane Science, November 24, 2000; 290(5496): 1567 - 1570. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
