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Proceedings Paper

mb-FLIM: model-based fluorescence lifetime imaging
Author(s): Qiaole Zhao; Ian Ted Young; Raymond Schouten; Sjoerd Stallinga; Kees Jalink; Sander de Jong
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Paper Abstract

We have developed a model-based, parallel procedure to estimate fluorescence lifetimes. Multiple frequencies are present in the excitation signal. Modeling the entire fluorescence and measurement process produces an analytical ratio of polynomials in the lifetime variable τ. A non-linear model-fitting procedure is then used to estimate τ. We have analyzed this model-based approach by simulating a 10 μM fluorescein solution (τ = 4 ns) and all relevant noise sources. We have used real LED data to drive the simulation. Using 240 μs of data, we estimate τ = 3.99 ns. Preliminary experiments on real fluorescent images taken from fluorescein solutions (measured τ = 4.1 ns), green plastic test slides (measured τ = 3.0 ns), and GFP in U2OS (osteosarcoma) cells (measured τ = 2.1 ns) demonstrate that this model-based measurement technique works.

Paper Details

Date Published: 9 February 2012
PDF: 11 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250M (9 February 2012); doi: 10.1117/12.905358
Show Author Affiliations
Qiaole Zhao, Technische Univ. Delft (Netherlands)
Ian Ted Young, Technische Univ. Delft (Netherlands)
Raymond Schouten, Technische Univ. Delft (Netherlands)
Sjoerd Stallinga, Technische Univ. Delft (Netherlands)
Kees Jalink, Netherlands Cancer Institute (Netherlands)
Sander de Jong, Lambert Instruments BV (Netherlands)

Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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