NADH Distribution in Live Progenitor Stem Cells by Phasor-Fluorescence Lifetime Image Microscopy
NADH Distribution in Live Progenitor Stem Cells by Phasor-Fluorescence Lifetime Image Microscopy
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DOI：
10.1016/j.bpj.2012.05.038
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发表时间：
2012-07-03
影响因子：
3.4
通讯作者：
Gratton, Enrico
中科院分区：
文献类型：
--
作者：
Wright, Belinda K.;Andrews, Laura M.;Gratton, Enrico
NADH 是一种与细胞呼吸相关的天然荧光代谢物。利用游离和结合 NADH 的不同荧光寿命有可能量化结合和游离 NADH 的相对量，从而增强对细胞过程（包括细胞凋亡、癌症病理学和酶动力学）的理解。我们使用相量荧光寿命图像显微镜方法对活的未分化和分化的成肌细胞的游离形式和结合形式的 NADH 进行空间图谱。相量方法以图形方式描绘了像素级寿命的变化，而不需要拟合衰减。通过血清饥饿诱导分化的细胞和未分化细胞的细胞核中 NADH 的空间分布的比较显示结合和游离 NADH 的不同分布。未分化细胞的寿命较短，表明细胞核中存在游离 NADH，而较长的寿命则归因于细胞核外存在结合的 NADH。分化细胞表现出游离 NADH 的重新分布，细胞核内游离 NADH 的相对浓度降低，而大部分 NADH 存在于细胞质中。
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.