Optical metabolic imaging of live tissue cultures

The fluorescence properties, both intensity and fluorescence lifetime, of NADH and FAD, two coenzymes of metabolism, are sensitive, high resolution measures of cellular metabolism. However, often in vivo measurements of tissue are not feasible. In this study, we investigate the stability over time of two-photon auto-fluorescence imaging of NADH and FAD in live-cultured tissues. Our results demonstrate that cultured tissues remain viable for at least several days post excision. Furthermore, the optical redox ratio, NADH fluorescence lifetime, and FAD fluorescence lifetime do not significantly change in the cultured tissues over time. With these findings, we demonstrate the potential of sustained tissue culture techniques for optical metabolic imaging.

[1]  Daniel A Fletcher,et al.  Tissue Geometry Determines Sites of Mammary Branching Morphogenesis in Organotypic Cultures , 2006, Science.

[2]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .

[3]  Melissa C. Skala,et al.  Optical imaging of metabolism in HER2 overexpressing breast cancer cells , 2011, Biomedical optics express.

[4]  B. Chance,et al.  Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. , 1979, The Journal of biological chemistry.

[5]  Jens Eickhoff,et al.  In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia. , 2007, Journal of biomedical optics.

[6]  O. Warburg.  On the origin of cancer cells. , 1956, Science.

[7]  W Cramer,et al.  On the Origin of Cancer* , 1938, British medical journal.

[8]  N. Ramanujam,et al.  In vivo multiphoton microscopy of NADH and FAD redox states, fluorescence lifetimes, and cellular morphology in precancerous epithelia , 2007, Proceedings of the National Academy of Sciences.

[9]  Alex J. Walsh,et al.  Ex vivo optical metabolic measurements from cultured tissue reflect in vivo tissue status , 2012, Journal of biomedical optics.

[10]  O. Warburg.  [Origin of cancer cells]. , 1956, Oncologia.

[11]  Ruth E. Cameron,et al.  A Multifunctional 3D Co-Culture System for Studies of Mammary Tissue Morphogenesis and Stem Cell Biology , 2011, PloS one.