Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy
Subdiffraction multicolor imaging of the nuclear periphery with 3D structured illumination microscopy
复制标题
DOI：
10.1126/science.1156947
复制
发表时间：
2008-06-06
期刊：
影响因子：
56.9
通讯作者：
Sedat, John W.
中科院分区：
文献类型：
--
作者：
Schermelleh, Lothar;Carlton, Peter M.;Sedat, John W.
荧光光学显微镜能够高特异性地对细胞成分进行多色可视化，但直到最近，其应用一直受到空间分辨率内在极限的限制。我们应用三维结构照明显微镜（3D - SIM）来规避这一限制并研究哺乳动物细胞核。通过同时对染色质、核纤层和核孔复合体（NPC）进行成像，我们观察到了一些常规显微镜无法检测到的特征。我们能够分辨出与核纤层网络中的通道以及周边异染色质共定位的单个核孔复合体。我们能够区分不同核孔复合体成分的定位，并检测到前期核膜的双层内陷，这在以前只有通过电子显微镜才能看到。多色3D - SIM为分析超出发射光衍射极限的亚细胞结构提供了新的、简便的可能性。
Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution. We applied three- dimensional structured illumination microscopy ( 3D- SIM) to circumvent this limit and to study the mammalian nucleus. By simultaneously imaging chromatin, nuclear lamina, and the nuclear pore complex ( NPC), we observed several features that escape detection by conventional microscopy. We could resolve single NPCs that colocalized with channels in the lamin network and peripheral heterochromatin. We could differentially localize distinct NPC components and detect double- layered invaginations of the nuclear envelope in prophase as previously seen only by electron microscopy. Multicolor 3D- SIM opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light.