Special Section on Visible Fluorescent Proteins

Nonlinear optical imaging and spectral-lifetime computational analysis of endogenous and exogenous fluorophores in breast cancer

[+] Author Affiliations
Paolo P. Provenzano

University of Wisconsin, Departments of Pharmacology and Biomedical Engineering and Laboratory for Optical and Computational Instrumentation and Paul P. Carbone Comprehensive Cancer Center, Madison, Wisconsin 53706

Curtis T. Rueden

University of Wisconsin, Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin 53706

Steve M. Trier

University of Wisconsin, Department of Biomedical Engineering and Laboratory for Optical and Computational Instrumentation and Paul P. Carbone Comprehensive Cancer Center, Madison, Wisconsin 53706

Long Yan

University of Wisconsin, Department of Biomedical Engineering and Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin 53706

Suzanne M. Ponik

University of Wisconsin, Departments of Pharmacology and Biomedical Engineering and Laboratory for Optical and Computational Instrumentation and Paul P. Carbone Comprehensive Cancer Center, Madison, Wisconsin 53706

David R. Inman

University of Wisconsin, Department of Pharmacology and Paul P. Carbone Comprehensive Cancer Center, Madison, Wisconsin 53706

Patricia J. Keely

University of Wisconsin, Departments of Pharmacology and Biomedical Engineering and Laboratory for Optical and Computational Instrumentation and Paul P. Carbone Comprehensive Cancer Center, Madison, Wisconsin 53706

Kevin W. Eliceiri

University of Wisconsin, Department of Biomedical Engineering and Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin 53706

J. Biomed. Opt. 13(3), 031220 (July 02, 2008). doi:10.1117/1.2940365
History: Received November 14, 2007; Revised November 18, 2008; Accepted November 18, 2008; Published July 02, 2008
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Multiphoton laser scanning microscopy (MPLSM) utilizing techniques such as multiphoton excitation (MPE), second harmonic generation (SHG), and multiphoton fluorescence lifetime imaging and spectral lifetime imaging (FLIM and SLIM, respectively) are greatly expanding the degree of information obtainable with optical imaging in biomedical research. The application of these nonlinear optical approaches to the study of breast cancer holds particular promise. These noninvasive, multidimensional techniques are well suited to image exogenous fluorophores that allow relevant questions regarding protein localization and signaling to be addressed both in vivo and in vitro. Furthermore, MPLSM imaging of endogenous signals from collagen and fluorophores such as nicotinamide adenine dinucleotide (NADH) or flavin adenine dinucleotide (FAD), address important questions regarding the tumor-stromal interaction and the physiologic state of the cell. We demonstrate the utility of multimodal MPE/SHG/FLIM for imaging both exogenous and/or endogenous fluorophores in mammary tumors or relevant 3-D systems. Using SLIM, we present a method for imaging and differentiating signals from multiple fluorophores that can have overlapping spectra via SLIM Plotter—a computational tool for visualizing and analyzing large spectral-lifetime data sets.

Figures in this Article
© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Paolo P. Provenzano ; Curtis T. Rueden ; Steve M. Trier ; Long Yan ; Suzanne M. Ponik, et al.
"Nonlinear optical imaging and spectral-lifetime computational analysis of endogenous and exogenous fluorophores in breast cancer", J. Biomed. Opt. 13(3), 031220 (July 02, 2008). ; http://dx.doi.org/10.1117/1.2940365


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