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DOI: 10.3791/64268-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This study describes a method for measuring absolute DNA densities within adherent cell nuclei, utilizing Voronoi tessellation of single-molecule localization microscopy (SMLM) data. This approach enables the investigation of spatial constraints in chromatin structures, linking genetic information with cell cycle stages.
本协议描述了一种使用单分子定位显微镜数据、已知体积、基因组大小和细胞周期阶段的Voronoi细分来测量贴壁细胞核内绝对DNA密度的方法。
本方法允许在细胞核中进行绝对DNA密度测量,以研究染色质结构中的空间限制,否则这些结构只能通过翻译后组蛋白修饰来表征。Voronoi 镶嵌与 SMLM 相结合,可以根据每平方微米碱基对 DNA 进行绝对密度估计。唯一需要的先验知识是被测细胞核的总 DNA 含量。
在未来的实验中,研究绝对密度差异是否与其他方法的生物信息相关,例如表观遗传修饰的免疫荧光或 Hi-C 数据,将会很有趣。首先通过将预先录制的单个图像平铺在一起来重建扫描区域。打开 CellProfiler,然后加载管道 cellcycleanalysis.cpproj。
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