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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 >Two-photon FLIM-FRET Microscopy for Protein Localization
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Two-photon FLIM-FRET Microscopy for Protein Localization

机译：两光子FLIM-FRET显微镜用于蛋白质定位

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摘要

Intensity based FRET visualizes protein molecules in living cells and tissues. Lifetime techniques, however, will demonstrate the dynamic functional activity of the protein molecules, because its signal does not depend on changes in fluorophore concentration or excitation intensity. If a laser pulse excites a large number of similar molecules with a similar local environment, and as long as no energy is transferred to another molecule, the lifetime is the "natural fluorescence lifetime". If energy is transferred, however, the actual fluorescence lifetime is less than the natural lifetime, because an additional path for de-excitation is present. With the occurrence of FRET, strong energy transfer results in extreme quenching of the donor fluorescence and a decrease in the fluorescence lifetime. In this paper we will explain the development of the two-photon FLIM-FRET microscopy with our existing multiphoton microscopy and demonstrate the change in donor lifetime by photobleaching the acceptor molecules in living cells.
机译：基于强度的FRET使活细胞和组织中的蛋白质分子可视化。但是，终生技术将证明蛋白质分子的动态功能活性，因为其信号不取决于荧光团浓度或激发强度的变化。如果激光脉冲激发具有相似局部环境的大量相似分子，并且只要没有能量转移到另一个分子，则该寿命就是“自然荧光寿命”。但是，如果转移了能量，则实际的荧光寿命会小于自然寿命，因为存在去激励的附加路径。随着FRET的发生，强烈的能量转移会导致供体荧光的极端淬灭并缩短荧光寿命。在本文中，我们将用我们现有的多光子显微镜解释两光子FLIM-FRET显微镜的发展，并通过光漂白活细胞中的受体分子来证明供体寿命的变化。

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