Fluorescence lifetime imaging microscopy (FLIM) has been demonstrated as advantageous at discriminating between free and protein-bound forms of the NADH coenzyme, providing not only with the lifetimes of the both states (shorter τ₁ and longer τ₂), but also with the relative concentrations of both (fractions a₁ and a₂ correspondingly). Given the role of NADH in cellular energetics, NADH FLIM has been applied for the noninvasive characterization of metabolic changes in a range of pathologies. However, for the discrimination of pathological states, a proper characterization of NADH fluorescence lifetime dynamics at physiological conditions has to be conducted. We have applied FLIM NADH for the characterization of metabolic changes during cell culture growth. Our results demonstrate that during the exponential growth stage there's a well expressed trends of gradual decrease of the free/bound ratio, as measured from the center from the cell colonies. At the same time the cells at the edges of a colony exhibit higher values of the ratio. Several possible reasons for the phenomena observed are discussed.

Date Published: 25 November 2009
PDF: 8 pages
Proc. SPIE 7634, Optical Sensors and Biophotonics, 76340P (25 November 2009); doi: 10.1117/12.855051

Published in SPIE Proceedings Vol. 7634:
Optical Sensors and Biophotonics
Xingde Li; Qingming Luo; Vasilis Ntziachristos; Yoshiaki Yasuno, Editor(s)