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首页> 外文会议>Conference on Multiphoton Microscopy in the Biomedical Sciences III Jan 26-28, 2003 San Jose, California, USA >Multiphoton microspectroscopy in living plant cells
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Multiphoton microspectroscopy in living plant cells

机译：活植物细胞中的多光子显微光谱

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摘要

Microspectroscopicxmeasurements in plant cells are complicated by the presence of dense cellular structures such as the cell wall that causes severe light scattering. In addition, the low penetration depth of the excitation light limits the fluorescence signal originating from deeper cell layers in thick multi-cellular plant preparations when single-photon excitation (SPE) is applied. However, two-photon excitation (TPE) can overcome these problems. We report on two-photon microscopy studies of Histone 2B-YFP, a nuclear-expressed protein involved in chromatin packaging. In contrast to SPE, TPE allows imaging throughout the whole root. Therefore by using TPE it was also possible to visualize the root quiescent centers using SCARECROW-EGFP localized in the middle of the root. The interactions between various members of the Arabidopsis thaliana embryogenesis receptor kinase family (AtSERK) have been studied by monitoring Foerster resonance energy transfer (FRET) between AtSERK-ECFP and -EYFP fusion proteins using fluorescence lifetime imaging microscopy (FLIM) of the two-photon excited ECFP component.
机译：由于存在致密的细胞结构（例如细胞壁）会导致严重的光散射，因此植物细胞中的显微光谱测量变得很复杂。另外，当施加单光子激发（SPE）时，激发光的低穿透深度限制了源自厚多细胞植物制剂中较深细胞层的荧光信号。但是，双光子激发（TPE）可以克服这些问题。我们报道了组蛋白2B-YFP的两光子显微镜研究，组蛋白2B-YFP是参与染色质包装的核表达蛋白。与SPE相比，TPE可以在整个根部成像。因此，通过使用TPE，还可以使用位于根中间的SCARECROW-EGFP可视化根静止中心。通过使用双光子的荧光寿命成像显微镜（FLIM）监视AtSERK-ECFP和-EYFP融合蛋白之间的Foerster共振能量转移（FRET），研究了拟南芥胚胎发生受体激酶家族（AtSERK）各个成员之间的相互作用。兴奋的ECFP组件。

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