Optical imaging of mitochondrial bioenergetics

BIOL 140

Julia Lyubovitsky, jlyubovi@uci.edu1, Joel A. Spencer, bogi@uci.edu2, Tatiana B. Krasieva, tkrasiev@uci.edu1, Chung-ho Sun1, Bogi Andersen, bogi@uci.edu2, and Bruce J. Tromberg, bjtrombe@uci.edu1. (1) Department of Surgery, Beckman Laser Institute, 1002 Health Sciences Road, Irvine, CA 92612, (2) Departments of Medicine and Biological Chemistry, UCI, Irvine, 92612
We characterized structural and metabolic heterogeneity in a mitochondrial distribution of ex-vivo normal, pathological mice corneas and isolated single cells using non-invasive two-photon ratiometric redox fluorometry method (TPRRF). The method is based on cellular fluorescence from reduced nicotinamide adenine dinucleotide (NADH) and oxidized flavin adenine dinucleotide (FAD). Employing NADH/FAD+ intensity ratio we detect higher metabolic activity in the endothelial layer of cornea as compared to an epithelial layer located further away from the metabolites. For the ACBT glioblastoma cells migrating in collagen and metrogel matrices the ratio is highly susceptible to noise and is observed to decrease by almost 50% in the optical sections with low quality of signal. Day to day uncertainty is ~20% for different cells. Thus, cellular behavior is complex and optical measurements on single cells may not be adequate to quantitatively describe cellular energetics. Therefore, we are measuring cellular metabolism in-vivo in a thick tissue such as cornea.
 

Chemistry and Metabolism
4:30 PM-6:30 PM, Tuesday, 12 September 2006 Moscone Center -- Hall D, Poster

Division of Biological Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006