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Quantitative Molecular Imaging in Living Cells via FLIM

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Reviews in Fluorescence 2010

Part of the book series: Reviews in Fluorescence ((RFLU,volume 2010))

Abstract

Fluorescence lifetime imaging microscopy (FLIM) employs fluorophore lifetime, rather than fluorescence intensity, for image contrast. Compared to intensity-based methods, lifetime imaging requires less calibration and/or correction for fluorophore concentration variations, photobleaching, and other artifacts that affect intensity measurements. We describe FLIM applications to probe the microenvironments of endogenous and exogenous fluorophores, including measurements of cellular metabolic co-factors, intracellular and extracellular oxygen, and molecular interactions via Förster resonance energy transfer (FRET). Several applications of FLIM for quantitative, live cell imaging are presented, including studies of cellular metabolic pathways, improved FRET detection of oncogene association, microfluidic bioreactor characterization for continuous cell culture, and improved analysis of FLIM images including image restoration and precision enhancement.

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Acknowledgments

We would like to acknowledge technical contributions from and helpful discussions with Drs. Mei Wu, Sofia D. Merajver, Dhruv Sud, Wei Zhong, Paul Urayama, David G. Beer, Jennifer Linderman, Shuichi Takayama, and Geeta Mehta, as well as Karl A. Merrick, Khamir Mehta, Jonathon Girroir, and Joe Delli. This work was supported in part by funding from the National Institutes of Health (CA-112173, CA-77612, and CA-114542) and The Whitaker Foundation.

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  1. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA

    Ching-Wei Chang & Mary-Ann Mycek

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  1. Ching-Wei Chang
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Correspondence to Mary-Ann Mycek .

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  1. Institute of Fluorescence, University of Maryland, E. Pratt St. 701, Baltimore, 21202, Maryland, USA

    Chris D. Geddes

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Chang, CW., Mycek, MA. (2012). Quantitative Molecular Imaging in Living Cells via FLIM. In: Geddes, C. (eds) Reviews in Fluorescence 2010. Reviews in Fluorescence, vol 2010. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-9828-6_8

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