A fast global fitting algorithm for fluorescence lifetime imaging microscopy based on image segmentation.

Details

Serval ID
serval:BIB_40CFE996C6DF
Type
Article: article from journal or magazin.
Collection
Publications
Title
A fast global fitting algorithm for fluorescence lifetime imaging microscopy based on image segmentation.
Journal
Biophysical Journal
Author(s)
Pelet S., Previte M.J.R., Laiho L.H., So P.T.
ISSN
0006-3495 (Print)
ISSN-L
0006-3495
Publication state
Published
Issued date
2004
Volume
87
Number
4
Pages
2807-2817
Language
english
Abstract
Global fitting algorithms have been shown to improve effectively the accuracy and precision of the analysis of fluorescence lifetime imaging microscopy data. Global analysis performs better than unconstrained data fitting when prior information exists, such as the spatial invariance of the lifetimes of individual fluorescent species. The highly coupled nature of global analysis often results in a significantly slower convergence of the data fitting algorithm as compared with unconstrained analysis. Convergence speed can be greatly accelerated by providing appropriate initial guesses. Realizing that the image morphology often correlates with fluorophore distribution, a global fitting algorithm has been developed to assign initial guesses throughout an image based on a segmentation analysis. This algorithm was tested on both simulated data sets and time-domain lifetime measurements. We have successfully measured fluorophore distribution in fibroblasts stained with Hoechst and calcein. This method further allows second harmonic generation from collagen and elastin autofluorescence to be differentiated in fluorescence lifetime imaging microscopy images of ex vivo human skin. On our experimental measurement, this algorithm increased convergence speed by over two orders of magnitude and achieved significantly better fits.
Keywords
Algorithms, Cells, Cultured, Collagen/metabolism, Collagen/ultrastructure, Fibroblasts/cytology, Fibroblasts/metabolism, Humans, Image Enhancement/methods, Image Interpretation, Computer-Assisted/methods, Microscopy, Fluorescence/methods, Numerical Analysis, Computer-Assisted, Reproducibility of Results, Sensitivity and Specificity, Skin/cytology, Skin/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
25/09/2012 14:38
Last modification date
20/08/2019 13:39
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