Fluorescence recovery after photobleaching: Application to nuclear proteins
Fluorescence recovery after photobleaching: Application to nuclear proteins
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DOI：
10.1007/b102214
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发表时间：
2005-01-01
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影响因子：
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通讯作者：
Houtsmuller, AB
中科院分区：
文献类型：
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作者：
Houtsmuller, AB
光漂白后的荧光再分布（FRAP）自被引入细胞生物学研究以来，受到越来越多的关注。该方法是在20世纪70年代开发的，当时其生物学应用主要集中在细胞膜的荧光标记成分的流动性上。20世纪80年代共聚焦扫描显微镜的发展促进了在没有专门设备的情况下对细胞内部分子行为的准确研究。然而，FRAP还没有像今天这样流行，可能是因为纯化和标记蛋白质或其他化合物以及将它们注射到细胞中所需的专用和耗时的方法。GFP技术的发展所带来的革命最终导致FRAP在研究活细胞中蛋白质行为方面的应用的巨大推动。最后，个人计算机的速度和内存的不断增加允许计算机模拟FRAP实验，用于分析复杂的3-D FRAP数据，并用于开发新的FRAP分析。在这里，我们讨论了几个变种的FRAP的基础上，它的应用调查的蛋白质在活细胞核中的行为。
Fluorescence redistribution after photobleaching (FRAP) has received increasing attention ever since it was first introduced into cell biological research. The method was developed in the 1970s, when its biological application mainly focused on the mobility of fluorescently labelled constituents of the cell membrane. The development of confocal scanning microscopy in the 1980s facilitated accurate investigation of the behaviour of molecules in the inside of cells without specialised equipment. However, FRAP did not yet become as popular as it is today, probably because of the dedicated and time-consuming methodology required to purify and label proteins or other compounds and, moreover, to inject them into cells. The revolution created by the development of GFP-technology finally lead to a tremendous boost of FRAP applications in studying the behaviour of proteins in the living cells. Finally, the ongoing increase of speed and memory of personal computers allows computer modelling of FRAP experiments for analysis of complex 3-D FRAP data, and for the development of new FRAP assays. Here we discuss several variants of FRAP on the basis of its application to the investigation of the behaviour of proteins in the living cell nucleus.