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Proceedings Paper

A hybrid global fitting algorithm for decay-associated images from fluorescence lifetime image microscopy data
Author(s): Aleksandr V. Smirnov; Christian A. Combs; Robert S Balaban; Kevin Tang; Jay R. Knutson
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Paper Abstract

Fluorescence lifetime imaging microscopy is a technique in which the fluorescence lifetime(s) of a fluorophore is measured at each spatially resolvable element of a microscope image. Imaging of fluorescence lifetimes enables biochemical reactions to be followed at each microscopically resolvable location within the cell. FLIM has thus become very useful for biomedical tissue imaging. Global analysis [1] is a method of recovering fluorescence decay parameters from either time-resolved emission spectra to yield Decay-Associated Spectra [2], or equivalently, from FLIM datasets to yield Decay-Associated Images. Global analysis offers a sensitive and non-invasive probe of metabolic state of intracellular molecules such as NADH. Using prior information, such as the spatial invariance of the lifetime of each fluorescent species in the image, to better refine the relevant parameters, global analysis can recover lifetimes and amplitudes more accurately than traditional pixel-by-pixel analysis. Here, we explain a method to analyze FLIM data so that more accurate lifetimes and DAIs can be computed in a reasonable time. This approach involves coupling an iterative global analysis with linear algebraic operations. It can be successfully applied to image, e.g. metabolic states of live cardiac myocytes, etc.

Paper Details

Date Published: 3 March 2009
PDF: 6 pages
Proc. SPIE 7184, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVI, 71841D (3 March 2009); doi: 10.1117/12.809844
Show Author Affiliations
Aleksandr V. Smirnov, National Institutes of Health (United States)
Christian A. Combs, National Institutes of Health (United States)
Robert S Balaban, National Institutes of Health (United States)
Kevin Tang, Yale Univ. (United States)
Jay R. Knutson, National Institutes of Health (United States)

Published in SPIE Proceedings Vol. 7184:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVI
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)

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