Yin Yandong, Rothenberg Eli
Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 550 1st Avenue, New York, NY 10016, USA.
Sci Rep. 2016 Aug 22;6:30819. doi: 10.1038/srep30819.
Super-resolution microscopy coupled with multiplexing techniques can resolve specific spatial arrangements of different components within molecular complexes. However, reliable quantification and analysis of such specific organization is extremely problematic because it is frequently obstructed by random co-localization incidents between crowded molecular species and the intrinsic heterogeneity of molecular complexes. To address this, we present a Triple-Pair-Correlation (TPC) analysis approach for unbiased interpretation of the spatial organization of molecular assemblies in crowded three-color super-resolution (SR) images. We validate this approach using simulated data, as well as SR images of DNA replication foci in human cells. This demonstrates the applicability of TPC in deciphering the specific spatial organization of molecular complexes hidden in dense multi-color super-resolution images.
超分辨率显微镜与多路复用技术相结合，可以解析分子复合物中不同组分的特定空间排列。然而，对这种特定组织进行可靠的量化和分析极具问题，因为它经常受到拥挤分子种类之间随机共定位事件以及分子复合物固有异质性的阻碍。为了解决这个问题，我们提出了一种三对关联（TPC）分析方法，用于无偏解释拥挤的三色超分辨率（SR）图像中分子组装体的空间组织。我们使用模拟数据以及人类细胞中DNA复制位点的SR图像对该方法进行了验证。这证明了TPC在解读隐藏于密集多色超分辨率图像中的分子复合物特定空间组织方面的适用性。
