Quantitative tomographic imaging of intermolecular FRET in small animals.
Quantitative tomographic imaging of intermolecular FRET in small animals.
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DOI：
10.1364/boe.3.003161
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发表时间：
2012-12-01
影响因子：
3.4
通讯作者：
Intes X
中科院分区：
文献类型：
--
作者：
Venugopal V;Chen J;Barroso M;Intes X
Forster共振能量转移(FRET)是两个荧光分子(给体和受体)在纳米范围内的非辐射能量转移。基于分子间FRET效率测量和FRET相互作用的化学计量测量作为感兴趣的定量参数，FRET成像方法已被应用于蛋白质组学研究和药物开发应用。重要的是，FRET在分子水平上提供了关于生物分子相互作用的信息，远远超出了标准显微镜技术的衍射极限。FRET在小动物成像中的应用将使生物医学研究人员能够在体内和原位研究发生在纳米范围内的生理过程。在这项工作中，描述了一种新的定量重建小动物FRET测量的方法，该方法将全场断层扫描采集系统与基于蒙特卡洛的分层重建方案相结合，并在小鼠模型中进行了验证。我们的主要目标是估计两种形式的供体分子的相对浓度，即参与FRETING的供体分子与邻近受体的供体分子和非FRETING供体分子的相对浓度。
Forster resonance energy transfer (FRET) is a nonradiative transfer of energy between two fluorescent molecules (a donor and an acceptor) in nanometer range proximity. FRET imaging methods have been applied to proteomic studies and drug discovery applications based on intermolecular FRET efficiency measurements and stoichiometric measurements of FRET interaction as quantitative parameters of interest. Importantly, FRET provides information about biomolecular interactions at a molecular level, well beyond the diffraction limits of standard microscopy techniques. The application of FRET to small animal imaging will allow biomedical researchers to investigate physiological processes occurring at nanometer range in vivo as well as in situ. In this work a new method for the quantitative reconstruction of FRET measurements in small animals, incorporating a full-field tomographic acquisition system with a Monte Carlo based hierarchical reconstruction scheme, is described and validated in murine models. Our main objective is to estimate the relative concentration of two forms of donor species, i.e., a donor molecule involved in FRETing to an acceptor close by and a nonFRETing donor molecule.