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Insight into cell metabolism will help interpret PETs, MRIs

short, intense laser pulses of harmless infrared instead of ultraviolet radiation. The technique, called multi-photon laser scanning microscopy (MPLSM), allowed the Vishwasrao team to measure NADH levels in cells with controlled levels of oxygen saturation without damaging the cells. And unlike other methods, such as PET and blood oxygen level dependent functional magnetic resonance imaging (BOLD-fMRI), MPLSM can simultaneously show how the orientation of NADH molecules changes (by measuring their anisotropy) within fractions of a nanosecond.

The results, said Vishwasrao, now a postdoctoral fellow at Columbia University, indicate that the unbound NADH molecules rotate much more quickly -- and therefore lose their fluorescence more quickly -- than bound NADH molecules.

"One bound NADH molecule is about as bright as 10 free ones," said Vishwasrao. "When we first got evidence that there was free NADH, we thought we made a big mistake. We thought we were crazy. We went back, and the more we talked about it, and the more experiments we did, it became clear. Other groups were seeing the same thing."

When the team used the data to calculate the proportion of bound-to-free NADH molecules in a section of tissue, they found that their calculations resolved inconsistencies that had troubled researchers for years. "The effect is large enough to account for the frequently seen problems," said Webb.

NADH is a good indicator of cell activity for several reasons. First, the molecule is ubiquitous in the mitochondria, where oxidative metabolism takes place. It also naturally fluoresces, which means it can be detected without adding artificial tracers or dyes. And because NADH is converted in the metabolic process to non-fluorescent NAD+, researchers can gauge how much oxidation is occurring in a cell based on its fluorescence.

With this new information, Vishwasrao said, scientists and physicians who study the effects of stroke, Alzheimer's disease and other
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Contact: Blaine P. Friedlander Jr.
bpf2@cornell.edu
607-254-8093
Cornell University News Service
11-Jul-2005


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