Gould Travis J, Gunewardene Mudalige S, Gudheti Manasa V, Verkhusha Vladislav V, Yin Shu-Rong, Gosse Julie A, Hess Samuel T
Department of Physics and Astronomy and Institute for Molecular Biophysics, 5709 Bennett Hall, University of Maine, Orono, Maine 04469, USA.
Nat Methods. 2008 Dec;5(12):1027-30. doi: 10.1038/nmeth.1271. Epub 2008 Nov 16.
Knowledge of the orientation of molecules within biological structures is crucial to understanding the mechanisms of cell function. We present a method to image simultaneously the positions and fluorescence anisotropies of large numbers of single molecules with nanometer lateral resolution within a sample. Based on a simple modification of fluorescence photoactivation localization microscopy (FPALM), polarization (P)-FPALM does not compromise speed or sensitivity. We show results for mouse fibroblasts expressing Dendra2-actin or Dendra2-hemagglutinin.
了解生物结构中分子的取向对于理解细胞功能机制至关重要。我们提出了一种方法,可在样品内以纳米横向分辨率同时成像大量单分子的位置和荧光各向异性。基于对荧光光激活定位显微镜(FPALM)的简单改进,偏振(P)-FPALM不会影响速度或灵敏度。我们展示了表达Dendra2-肌动蛋白或Dendra2-血凝素的小鼠成纤维细胞的结果。