The aim of this er vito study was to explore the potential of using the fluorescence lifetime of intraellular reduced nicotinamide adenine dinucleotide(phosphate)(NAD(P)H)as a label-free indicator to characterize the ...The aim of this er vito study was to explore the potential of using the fluorescence lifetime of intraellular reduced nicotinamide adenine dinucleotide(phosphate)(NAD(P)H)as a label-free indicator to characterize the di ferencs between human leukemic myeloid cells and normal mononuclear cells(MNC).The steady-state and time-resolved autofuorescence of two human leukemic myeloid cell lines(K562,HL60)and MNC were measured by a spectrofuorimeter.According to excitation-enmission matrix(EEM)analysis,the optimal emission of NAD(P)H in these cells suspensions occurred at 445 nm.Furthermore,the fuorescence lifetimes of NAD(P)H in leukemic myeloid cells and MNC were determined by fitting the time-resolved autofuorescence data.The mean fuorescence lifetimes of NAD(P)H in K562,HL60,and MNC cells were 557±1.19,4.45±0.71,and 7.31±0.60 ns,respectively.There was a significant diference in the mean lifetime of NAD(P)H between leukemic myeloid cells and MNC(p<0.05).The difference was essentally caused by the change in relative concentration of free and protein-bound NAD(P)H.This study suggests that the mean fuorescence lifetime of NAD(P)H might be a potential label-free indicator for differentiating leukemic myeloid cells from MNC.展开更多
基金supported by the National Natural Science Foundation of China(61275216)the Fujian Provincial Natural Science Foundation(2011J06022)+1 种基金the Science Research foundation of Ministry of Health&United Fujian Provincial Health and Education Project for Tackling the Key Research(WKJ2008-2-049)the Program for Changjiang Scholars and Innovative Research Team in University(IRT1115).
文摘The aim of this er vito study was to explore the potential of using the fluorescence lifetime of intraellular reduced nicotinamide adenine dinucleotide(phosphate)(NAD(P)H)as a label-free indicator to characterize the di ferencs between human leukemic myeloid cells and normal mononuclear cells(MNC).The steady-state and time-resolved autofuorescence of two human leukemic myeloid cell lines(K562,HL60)and MNC were measured by a spectrofuorimeter.According to excitation-enmission matrix(EEM)analysis,the optimal emission of NAD(P)H in these cells suspensions occurred at 445 nm.Furthermore,the fuorescence lifetimes of NAD(P)H in leukemic myeloid cells and MNC were determined by fitting the time-resolved autofuorescence data.The mean fuorescence lifetimes of NAD(P)H in K562,HL60,and MNC cells were 557±1.19,4.45±0.71,and 7.31±0.60 ns,respectively.There was a significant diference in the mean lifetime of NAD(P)H between leukemic myeloid cells and MNC(p<0.05).The difference was essentally caused by the change in relative concentration of free and protein-bound NAD(P)H.This study suggests that the mean fuorescence lifetime of NAD(P)H might be a potential label-free indicator for differentiating leukemic myeloid cells from MNC.
