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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 9 — Sep. 1, 2012
  • pp: 2244–2262

Application of non-negative matrix factorization to multispectral FLIM data analysis

Paritosh Pande, Brian E. Applegate, and Javier A. Jo  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 9, pp. 2244-2262 (2012)

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Existing methods of interpreting fluorescence lifetime imaging microscopy (FLIM) images are based on comparing the intensity and lifetime values at each pixel with those of known fluorophores. This method becomes unwieldy and subjective in many practical applications where there are several fluorescing species contributing to the bulk fluorescence signal, and even more so in the case of multispectral FLIM. Non-negative matrix factorization (NMF) is a multivariate data analysis technique aimed at extracting non-negative signatures of pure components and their non-negative abundances from an additive mixture of those components. In this paper, we present the application of NMF to multispectral time-domain FLIM data to obtain a new set of FLIM features (relative abundance of constituent fluorophores). These features are more intuitive and easier to interpret than the standard fluorescence intensity and lifetime values. The proposed approach, unlike several FLIM data analysis methods, is not limited by the number of constituent fluorescing species or their possibly complex decay dynamics. Moreover, the new set of FLIM features can be obtained by processing raw multispectral FLIM intensity data, thereby rendering time deconvolution unnecessary and resulting in lesser computational time and relaxed SNR requirements. The performance of the NMF method was validated on simulated and experimental multispectral time-domain FLIM data. The NMF features were also compared against the standard intensity and lifetime features, in terms of their ability to discriminate between different types of atherosclerotic plaques.

© 2012 OSA

OCIS Codes
(070.5010) Fourier optics and signal processing : Pattern recognition
(170.1580) Medical optics and biotechnology : Chemometrics
(170.1610) Medical optics and biotechnology : Clinical applications
(170.6920) Medical optics and biotechnology : Time-resolved imaging
(300.2530) Spectroscopy : Fluorescence, laser-induced
(170.6935) Medical optics and biotechnology : Tissue characterization

ToC Category:
Image Processing

Original Manuscript: July 6, 2012
Revised Manuscript: August 19, 2012
Manuscript Accepted: August 20, 2012
Published: August 24, 2012

Paritosh Pande, Brian E. Applegate, and Javier A. Jo, "Application of non-negative matrix factorization to multispectral FLIM data analysis," Biomed. Opt. Express 3, 2244-2262 (2012)

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