Multiphoton fluorescence imaging of NADH to quantify metabolic changes in epileptic tissue in vitro

Thomas H. Chia; Joseph Zinter; Dennis D. Spencer M.D.; Anne Williamson; Michael J. Levene

doi:10.1117/12.696928

12 February 2007 Multiphoton fluorescence imaging of NADH to quantify metabolic changes in epileptic tissue in vitro

Thomas H. Chia, Joseph Zinter, Dennis D. Spencer M.D., Anne Williamson, Michael J. Levene

Proceedings Volume 6442, Multiphoton Microscopy in the Biomedical Sciences VII; 64421P (2007) https://doi.org/10.1117/12.696928
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

Citation Download Citation

Thomas H. Chia, Joseph Zinter, Dennis D. Spencer M.D., Anne Williamson, and Michael J. Levene "Multiphoton fluorescence imaging of NADH to quantify metabolic changes in epileptic tissue in vitro", Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64421P (12 February 2007); https://doi.org/10.1117/12.696928

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