| Literature DB >> 22828352 |
Belinda K Wright, Laura M Andrews, Julie Markham, Mark R Jones, Chiara Stringari, Michelle A Digman, Enrico Gratton.
Abstract
NADH is a naturally fluorescent metabolite associated with cellular respiration. Exploiting the different fluorescence lifetime of free and bound NADH has the potential to quantify the relative amount of bound and free NADH, enhancing understanding of cellular processes including apoptosis, cancer pathology, and enzyme kinetics. We use the phasor-fluorescence lifetime image microscopy approach to spatially map NADH in both the free and bound forms of live undifferentiated and differentiated myoblast cells. The phasor approach graphically depicts the change in lifetime at a pixel level without the requirement for fitting the decay. Comparison of the spatial distribution of NADH in the nucleus of cells induced to differentiate through serum starvation and undifferentiated cells show differing distributions of bound and free NADH. Undifferentiated cells displayed a short lifetime indicative of free NADH in the nucleus and a longer lifetime attributed to the presence of bound NADH outside of the nucleus. Differentiating cells displayed redistribution of free NADH with decreased relative concentration of free NADH within the nucleus whereas the majority of NADH was found in the cytoplasm.Entities:
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Year: 2012 PMID： 22828352 PMCID： PMC3388207 DOI： 10.1016/j.bpj.2012.05.038
Source DB: PubMed Journal: Biophys J ISSN： 0006-3495 Impact factor: 4.033