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Phasor‐flim analysis of NADH distribution and localization in the nucleus of live progenitor myoblast cells
Author(s) -
Wright Belinda K.,
Andrews Laura M.,
Jones Mark R.,
Stringari Chiara,
Digman Michelle A.,
Gratton Enrico
Publication year - 2012
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.22121
Subject(s) - nucleus , microbiology and biotechnology , progenitor cell , nuclear localization sequence , phasor , cofactor , cell , biology , cell nucleus , chemistry , biochemistry , stem cell , enzyme , power (physics) , physics , electric power system , quantum mechanics
Analysis of the cellular distributions of coenzymes including NADH may aid in understanding a cells metabolic status. We altered serum concentration (0, 2, and 10%) to induce living myoblast cells to undergo the early stages of differentiation. Through microscopy and phasor‐FLIM, we spatially mapped and identified variations in the distribution of free and bound NADH. Undifferentiated cells displayed abundant free NADH within the nucleus along with specific regions of more bound NADH. Complete serum starvation dramatically increased the fraction of bound NADH in the nucleus, indicating heightened requirement for transcriptional processes. In comparison, cells exposed to 2% serum exhibited intermediate free nuclear NADH fraction. Overall our results suggest an order of events in which a cell metabolic status alters significantly during the early stages of serum induced differentiation. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.