Imaging intracellular fluorescent proteins at nanometer resolution
Imaging intracellular fluorescent proteins at nanometer resolution
复制标题
DOI：
10.1126/science.1127344
复制
发表时间：
2006-09-15
期刊：
影响因子：
56.9
通讯作者：
Hess, Harald F.
中科院分区：
文献类型：
--
作者：
Betzig, Eric;Patterson, George H.;Hess, Harald F.
我们介绍一种以纳米空间分辨率对细胞内蛋白质进行光学成像的方法。许多可光激活荧光蛋白分子的稀疏子集被激活、定位（精度约为2到25纳米），然后漂白。来自所有子集的总体位置信息随后被组合成一幅超高分辨率图像。我们利用这种被称为光激活定位显微镜的方法，对溶酶体和线粒体薄片中的特定目标蛋白质进行成像；在固定的完整细胞中，我们对粘着斑处的纽蛋白、片足内的肌动蛋白以及逆转录病毒蛋白Gag在质膜上的分布进行了成像。
We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized ( to similar to 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method - termed photoactivated localization microscopy - to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.