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Proceedings Paper

Multiphoton fluorescence imaging of NADH to quantify metabolic changes in epileptic tissue in vitro
Author(s): Thomas H. Chia; Joseph Zinter; Dennis D. Spencer; Anne Williamson; Michael J. Levene
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Paper Abstract

A powerful advantage of multiphoton microscopy is its ability to image endogenous fluorophores such as the ubiquitous coenzyme NADH in discrete cellular populations. NADH is integral in both oxidative and non-oxidative cellular metabolism. NADH loses fluorescence upon oxidation to NAD+; thus changes in NADH fluorescence can be used to monitor metabolism. Recent studies have suggested that hypo metabolic astrocytes play an important role in cases of temporal lobe epilepsy (TLE). Current theories suggest this may be due to defective and/or a reduced number of mitochondria or dysfunction of the neuronal-astrocytic metabolic coupling. Measuring NADH fluorescence changes following chemical stimulation enables the quantification of the cellular distribution of metabolic anomalies in epileptic brain tissue compared to healthy tissue. We present what we believe to be the first multiphoton microscopy images of NADH from the human brain. We also present images of NADH fluorescence from the hippocampus of the kainate-treated rat TLE model. In some experiments, human and rat astrocytes were selectively labeled with the fluorescent dye sulforhodamine 101 (SR101). Our results demonstrate that multiphoton microscopy is a powerful tool for assaying the metabolic pathologies associated with temporal lobe epilepsy in humans and in rodent models.

Paper Details

Date Published: 12 February 2007
PDF: 5 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64421P (12 February 2007); doi: 10.1117/12.696928
Show Author Affiliations
Thomas H. Chia, Yale Univ. (United States)
Joseph Zinter, Yale Univ. (United States)
Dennis D. Spencer, Yale Univ. (United States)
Anne Williamson, Yale Univ. (United States)
Michael J. Levene, Yale Univ. (United States)

Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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