School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, United States.
ACS Nano. 2012 Sep 25;6(9):7928-34. doi: 10.1021/nn3023624. Epub 2012 Aug 1.
Individual chromatin molecules contain valuable genetic and epigenetic information. To date, there have not been reliable techniques available for the controlled stretching and manipulation of individual chromatin fragments for high-resolution imaging and analysis of these molecules. We report the controlled stretching of single chromatin fragments extracted from two different cancerous cell types (M091 and HeLa) characterized through fluorescence microscopy and atomic force microscopy (AFM). Our method combines soft lithography with molecular stretching to form ordered arrays of more than 250,000 individual chromatin fragments immobilized into a beads-on-a-string structure on a solid transparent support. Using fluorescence microscopy and AFM, we verified the presence of histone proteins after the stretching and transfer process.
单个染色质分子包含有价值的遗传和表观遗传信息。迄今为止,还没有可靠的技术可用于对单个染色质片段进行受控拉伸和操作,以实现对这些分子的高分辨率成像和分析。我们报告了通过荧光显微镜和原子力显微镜(AFM)对从两种不同癌细胞类型(M091 和 HeLa)中提取的单个染色质片段进行的受控拉伸。我们的方法将软光刻与分子拉伸相结合,在透明固体支撑物上形成固定在珠串结构中的超过 250000 个单个染色质片段的有序阵列。通过荧光显微镜和 AFM,我们在拉伸和转移过程后验证了组蛋白蛋白的存在。