Measurement of Mitochondrial Membrane Potential in Leukocyte Suspension by Fluorescent Spectroscopy
Measurement of Mitochondrial Membrane Potential in Leukocyte Suspension by Fluorescent Spectroscopy
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DOI：
10.1007/s10517-014-2547-4
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发表时间：
2014-06-01
影响因子：
0.7
通讯作者：
Kondrat'eva, E. V.
中科院分区：
文献类型：
--
作者：
Lobanova, E. G.;Kondrat'eva, E. V.
本文介绍了一种利用灵敏的荧光阳离子活性染料5，5 '，6，6'-四氯-1，1 '，3，3'-四乙基苯并咪唑碳菁（JC-1）和分光光度法测定分离的白细胞悬液中线粒体膜电位的方法。JC-1单体迅速穿透活细胞的线粒体膜并形成以红色荧光（λ = 585 nm）为特征的聚集体。在线粒体膜去极化（细胞凋亡的早期迹象）的情况下，JC-1不在线粒体中积累，并且作为以绿色光谱荧光（λ = 510 nm）为特征的单体存在于细胞质中。该方法可用于通过评估线粒体膜电位来评估活细胞的功能和调节能量代谢的机制。
A method for evaluating mitochondrial membrane potential in isolated leukocyte suspension with the use of sensitive fl uorescent cation-active dye 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine (JC-1) and spectrophotometry is described. JC-1 monomer rapidly penetrates through mitochondrial membrane of living cell and form aggregations characterized by red fl uorescence (lambda = 585 nm). In case of mitochondrial membrane depolarization (early sign of apoptosis), JC-1 is not accumulated in the mitochondria and is present in the cytoplasm as a monomer characterized by green spectral fl uorescence (lambda = 510 nm). The method can be used for evaluation of the function of living cells and mechanisms regulating energy metabolism by evaluating the mitochondrial membrane potential.