Howard Hughes Medical Institute, Janelia Farm Research Campus, 19700 Helix Drive, Ashburn, VA 20147-2408, USA.
Methods. 2010 Aug;51(4):458-63. doi: 10.1016/j.ymeth.2010.01.001. Epub 2010 Jan 12.
Mitochondria are difficult targets for microscopy because of their small size and highly compartmentalized, membranous interior. Super-resolution fluorescence microscopy methods have recently been developed that exceed the historical limits of optical imaging. Here we outline considerations and techniques in preparing to image the relative location of mitochondrial proteins using photoactivated localization microscopy (PALM). PALM and similar methods have the capacity to dramatically increase our ability to image proteins within mitochondria, and to expand our knowledge of the location of macromolecules beyond the current limits of immunoEM.
线粒体由于其体积小且内部高度分隔的膜结构,成为了显微镜观察的难点。最近开发的超分辨率荧光显微镜方法已经超越了传统光学成像的限制。本文概述了使用光激活定位显微镜(PALM)对线粒体蛋白相对位置进行成像的准备工作中的注意事项和技术。PALM 及类似方法有能力极大地提高我们在线粒体中对蛋白质成像的能力,并扩展我们对大分子位置的了解,超越免疫电子显微镜目前的限制。