Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET).
Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET).
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DOI：
10.1042/bst0311020
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发表时间：
2003-10
影响因子：
3.9
通讯作者：
D. Lidke;Peter Nagy;B. Barisas;Rainer Heintzmann;Janine N. Post;K. Lidke;A. H. Clayton;D. Arndt-Jovin;T. Jovin
中科院分区：
文献类型：
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作者：
D. Lidke;Peter Nagy;B. Barisas;Rainer Heintzmann;Janine N. Post;K. Lidke;A. H. Clayton;D. Arndt-Jovin;T. Jovin
我们报告了荧光偏振测量的实施和开发，以各向异性荧光寿命成像显微镜（rFLIM）和能量迁移Förster共振能量转移（emFRET）模式的形式，用于宽视场，共聚焦激光扫描显微镜和细胞流式细胞术。这些方法允许在体内评估细胞蛋白的旋转运动、关联和接近。它们特别适用于由可见荧光蛋白融合产生的探针，例如参与生长因子介导的信号转导的erbB受体酪氨酸激酶的研究。
We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow cytometry of cells. These methods permit the assessment of rotational motion, association and proximity of cellular proteins in vivo. They are particularly applicable to probes generated by fusions of visible fluorescence proteins, as exemplified by studies of the erbB receptor tyrosine kinases involved in growth-factor-mediated signal transduction.