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Methods Inf Med 2010; 49(05): 531-536
DOI: 10.3414/ME09-02-0046
Special Topic – Original Articles
Schattauer GmbH

Analysis of Fluorescence Lifetime Imaging Microscopy (FLIM) Data

Based on a Fully Automated Laguerre Deconvolution Method
P. Pande
1   Department of Biomedical Engineering, Texas A&M University, College Station, Texas, USA
,
C. A. Trivedi
1   Department of Biomedical Engineering, Texas A&M University, College Station, Texas, USA
,
J. A. Jo
1   Department of Biomedical Engineering, Texas A&M University, College Station, Texas, USA
› Author Affiliations
Further Information

Publication History

received: 10 October 2009

accepted: 23 January 2010

Publication Date:
17 January 2018 (online)

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Summary

Objectives: A novel Fluorescence Lifetime Imaging Microscopy (FLIM) deconvolution method based on the linear expansion of fluorescence decays on a set of orthonormal Laguerre functions was recently proposed. The Laguerre deconvolution method applies linear least-square estimation to estimate the expansion coefficients of all pixel decays simultaneously, performing at least two orders of magnitude faster than the other algorithms. In the original Laguerre FLIM deconvolution implementation, however, the Laguerre parameter α is selected using a heuristic approach, making it unsuitable for online applications.

Methods: In this study, we present a fully automated implementation of the Laguerre FLIM deconvolution, whereby the Laguerre parameter α is treated as a free parameter within a nonlinear least-squares optimization scheme.

Results: The performance of this method has been successfully validated on simulated data, and experimental FLIM images of standard fluorescent dyes and endogenous tissue fluorescence.

Conclusions: The main advantage of the proposed method is that it does not require any user intervention for tuning up the deconvolution process. Thus, we believe this method will facilitate the translation of FLIM to online applications, including real-time clinical diagnosis.

Keywords

Fluorescence Lifetime Imaging Microscopy - Laguerre deconvolution - nonlinear least squares - optimization
 

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