基于FRET荧光蛋白探针的活体定量分子影像方法探究

发布时间：2018-03-25 22:39

本文选题：荧光共振能量转移　切入点：基因编码FRET探针　出处：《北京生物医学工程》2016年06期

【摘要】：目的荧光共振能量转移(fluorescent resonance energy transfer,FRET)探针由于自身仍存在诸多局限,其在活体成像领域尚未得到广泛应用。本文旨在解决FRET探针在活体定量分子影像应用中所存在的瓶颈问题,即如何利用FRET探针精准计算目标物浓度,以及如何基于FRET探针实现三维成像。方法基于探针和目标物结合时的化学反应平衡关系,提出一种新型分析方法,以精确定量待测物[钙调素(calmodulin,Ca M)]的浓度。同时对于FRET探针的活体分子成像,结合荧光透射成像(3DFMT)方法,建立了可进行三维时空动态FRET检测的平台系统,对FRET探针的测量结果(即待测物浓度)进行实时定量的三维重建。结果准确地定量了待测物Ca M浓度,并得到了高质量的3D-FRET分布影像。结论提出FRET探针在活体成像应用领域两个关键问题的解决方案,为基因编码FRET探针向活体分子影像领域的推广奠定了重要的技术基础。
[Abstract]:The purpose of fluorescence resonance energy transfer (fluorescent resonance energy transfer, FRET) probe because of its own there are still many limitations, it has not been widely applied in the field of imaging. This paper aims to solve the bottleneck problem which exists in the FRET probe in vivo quantitative molecular imaging applications, namely how to use the FRET probe for accurate calculation of the target concentration, and based on FRET probe to realize 3D imaging. Methods based on the chemical reaction equilibrium relationship between probe and target combination, put forward a new analysis method, in order to precisely quantify the analytes [calmodulin (calmodulin Ca M)] concentration. At the same time for in vivo molecular FRET probe combined with fluorescence imaging, transmission imaging (3DFMT) method. To establish the platform system for dynamic 3D spatio-temporal FRET detection. The measurement results of FRET probe (i.e. analyte concentration) for real-time 3D reconstruction. The quasi The Ca M accurately quantitative concentration to be tested, and obtained the distribution of 3D-FRET image with high quality. The solution concludes two key problems of FRET probe in vivo imaging applications, lay the technical foundation for the promotion of important genes encoding FRET imaging to probe in vivo molecular field.

【作者单位】：清华大学医学院生物医学工程系;清华大学生命科学学院;
【基金】：国家自然科学基金(31370822,81171382,81371604) 北京市自然科学基金(7142089)资助
【分类号】：TP212.3;R318

【相似文献】