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首页> 外文会议>Conference on Multiphoton Microscopy in the Biomedical Sciences IV; 20040125-20040127; San Jose,CA; US >Dynamic and static fluorescence anisotropy in biological microscopy (rFLIM and em FRET)
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Dynamic and static fluorescence anisotropy in biological microscopy (rFLIM and em FRET)

机译：生物显微镜中的动态和静态荧光各向异性（rFLIM和em FRET）

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摘要

Fluorescence anisotropy, a measure of the polarization state of fluorescence emission, is a sensitive measure of molecular rotational motion and of resonance energy transfer (RET). We report here the formalism and application of dynamic and static fluorescence anisotropy measurements primarily intended for implementation in imaging systems. These include confocal laser scanning microscopes (CLSM) as well as wide-field instruments, in the latter case adapted for anisotropy-based dynamic frequency domain fluorescence lifetime imaging microscopy (FLIM), a method we denote as rFLIM. Anisotropy RET is one of the modalities used for fluorescence RET (FRET) determinations of the association, and proximity of cellular proteins in vivo. A requirement is the existence of intrinsic or extrinsic probes exhibiting homotransfer FRET (in our nomenclature, energy migration or emFRET) between like fluorophores. This phenomenon is particularly useful in studies of the activation and processing of transmembrane receptor tyrosine kinases involved in signal transduction and expressed as fusions with Visible Fluorescence Proteins (VFPs).
机译：荧光各向异性是荧光发射极化状态的一种度量，它是分子旋转运动和共振能量转移（RET）的敏感度量。我们在这里报告主要用于成像系统的动态和静态荧光各向异性测量的形式主义和应用。这些工具包括共聚焦激光扫描显微镜（CLSM）以及宽视野仪器，在后一种情况下，该仪器适用于基于各向异性的动态频域荧光寿命成像显微镜（FLIM），我们将其称为rFLIM。各向异性RET是用于荧光RET（FRET）测定体内细胞蛋白的缔合和接近性的方法之一。要求是存在内在或外在探针，它们在类似的荧光团之间表现出均相转移FRET（在我们的术语中，指的是能量迁移或emFRET）。这种现象在研究跨膜受体酪氨酸激酶的活化和加工过程中特别有用，该酪氨酸激酶参与信号转导并表达为与可见荧光蛋白（VFP）融合。

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