Measurement of replication structures at the nanometer scale using super-resolution light microscopy.

Measurement of replication structures at the nanometer scale using super-resolution light microscopy.
复制标题

使用超分辨率光显微镜在纳米尺度上测量复制结构。

DOI:
10.1093/nar/gkp901
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发表时间:
2010-01
影响因子:
14.9
通讯作者:
Cardoso MC
Cardoso MC
中科院分区:
生物学2区
文献类型:
--
作者:
Baddeley D;Chagin VO;Schermelleh L;Martin S;Pombo A;Carlton PM;Gahl A;Domaing P;Birk U;Leonhardt H;Cremer C;Cardoso MC

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DNA 复制与其他细胞过程类似,发生在动态大分子结构内。任何全面的理解最终都需要定量数据来建立和测试基因组复制模型。我们使用两种不同的超分辨率光学显微镜技术来直接测量和比较哺乳动物细胞中复制灶的大小和数量。该分析表明复制焦点的大小从 210 nm 到 40 nm 不等。值得注意的是,空间调制照明 (SMI) 和 3D 结构照明显微镜 (3D-SIM) 均显示平均大小为 125 nm,在整个 S 期中保守且与标记方法无关,表明基因组复制的基本单位。有趣的是,改进的光学 3D 分辨率识别出的复制焦点比之前报道的要多 3 至 5 倍。这些结果表明,光学纳米显微镜技术能够以以前只能通过电子显微镜达到的水平精确测量细胞结构,并强调高通量、多光谱 3D 分析的可能性。
DNA replication, similar to other cellular processes, occurs within dynamic macromolecular structures. Any comprehensive understanding ultimately requires quantitative data to establish and test models of genome duplication. We used two different super-resolution light microscopy techniques to directly measure and compare the size and numbers of replication foci in mammalian cells. This analysis showed that replication foci vary in size from 210 nm down to 40 nm. Remarkably, spatially modulated illumination (SMI) and 3D-structured illumination microscopy (3D-SIM) both showed an average size of 125 nm that was conserved throughout S-phase and independent of the labeling method, suggesting a basic unit of genome duplication. Interestingly, the improved optical 3D resolution identified 3- to 5-fold more distinct replication foci than previously reported. These results show that optical nanoscopy techniques enable accurate measurements of cellular structures at a level previously achieved only by electron microscopy and highlight the possibility of high-throughput, multispectral 3D analyses.
DOI: 10.2741/3600
发表时间: 2009-06-01
影响因子: 3.1
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