Use of spectral fluorescence resonance energy transfer to detect nitric oxide-based signaling events in isolated perfused lung.
Use of spectral fluorescence resonance energy transfer to detect nitric oxide-based signaling events in isolated perfused lung.
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使用光谱荧光共振能量转移来检测离体灌注肺中基于一氧化氮的信号事件。
DOI：
10.1002/0471142956.cy1213s45
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发表时间：
2008
影响因子：
--
通讯作者：
Bauer,EileenM
中科院分区：
文献类型：
--
作者：
StCroix,ClaudetteM;Bauer,EileenM
荧光共振能量转移（FRET）是一种适用于活细胞的荧光显微镜技术，当蛋白质与适当的供体和受体荧光团缀合时，能够检测单个蛋白质构象状态的变化或两个相互作用蛋白质之间的距离。基于共焦的光谱检测系统通过为图像的每个体素提供完整的光谱信息而无需切换滤光片，从而实现荧光图像的分辨率。此外，使用校准光谱，可以明确地分离重叠的供体和受体发射之间的串扰。本单元描述了使用基于共聚焦的光谱成像的一氧化氮（NO）敏感的FRET报告在完整的血管系统，隔离灌注小鼠肺。这种类型的原位成像方法允许在完整的活器官的适当生理微环境内可视化和研究时间分子信号传导事件。议定书细胞。45：12.13.1 - 12.13.12.© 2008年由约翰威利父子公司。
Fluorescence resonance energy transfer (FRET) is a fluorescence microscopy technique suitable for live cells and capable of detecting changes in the conformational state of a single protein or the distance between two interacting proteins when the proteins are conjugated with appropriate donor and acceptor fluorophores. Confocal‐based spectral detection systems enable the resolution of fluorescent images by providing full spectral information for each voxel of the image without switching of optical filters. Furthermore, using calibration spectra, it is possible to unambiguously separate the cross‐talk between overlapping donor and acceptor emissions. This unit describes the use of confocal‐based spectral imaging of nitric oxide (NO) sensitive FRET reporters in the vasculature of the intact, isolated perfused mouse lung. This type of in situ imaging approach allows the visualization and study of temporal molecular signaling events within the appropriate physiologic microenvironment of the intact, living organ.Curr. Protocol. Cytom. 45:12.13.1‐12.13.12. © 2008 by John Wiley & Sons, Inc.