A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells.
A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells.
复制标题
DOI：
10.1016/j.xpro.2021.100865
复制
发表时间：
2021-12-17
期刊：
影响因子：
--
通讯作者：
Cosma MP
中科院分区：
文献类型：
--
作者：
Martin L;Vicario C;Castells-García Á;Lakadamyali M;Neguembor MV;Cosma MP
在这里，我们描述了三种互补的基于显微镜的方法来量化培养贴壁细胞中染色质组织的形态变化：DNA变异系数的分析、无DNA核区域的测量以及核边缘染色质相关蛋白的量化。这些方法依赖于共焦成像和随机光学重建显微镜，可以快速、稳健地量化染色质压实。这些方法避免了成像条件之间的相互差异，并适用于每种类型的贴壁细胞。有关该协议的使用和执行的完整详细信息，请参阅。测量染色质压实的三种互补技术的协议 从共焦和超分辨率图像中量化染色质压实 CV 和 Black Space 分析核 DNA 分布异质性的评分 边缘分析测量核边缘周围蛋白质信号的富集 在这里，我们描述了三种基于互补显微镜的方法来量化培养贴壁细胞中染色质组织的形态变化：DNA 变异系数的分析、无 DNA 核区域的测量以及核边缘的染色质相关蛋白。这些方法依赖于共焦成像和随机光学重建显微镜，可以快速、稳健地量化染色质压实。这些方法避免了成像条件之间的相互差异，并适用于每种类型的贴壁细胞。
Here, we describe three complementary microscopy-based approaches to quantify morphological changes of chromatin organization in cultured adherent cells: the analysis of the coefficient of variation of DNA, the measurement of DNA-free nuclear areas, and the quantification of chromatin-associated proteins at the nuclear edge. These approaches rely on confocal imaging and stochastic optical reconstruction microscopy and allow a fast and robust quantification of chromatin compaction. These approaches circumvent inter-variability between imaging conditions and apply to every type of adherent cells. For complete details on the use and execution of this protocol, please refer to. Protocol for three complementary techniques to measure chromatin compaction Chromatin compaction is quantified from confocal and super-resolution images CV and Black Space analyses score for heterogeneity of nuclear DNA distribution Edge analysis measures the enrichment of a protein signal around the nuclear edge Here, we describe three complementary microscopy-based approaches to quantify morphological changes of chromatin organization in cultured adherent cells: the analysis of the coefficient of variation of DNA, the measurement of DNA-free nuclear areas, and the quantification of chromatin-associated proteins at the nuclear edge. These approaches rely on confocal imaging and stochastic optical reconstruction microscopy and allow a fast and robust quantification of chromatin compaction. These approaches circumvent inter-variability between imaging conditions and apply to every type of adherent cells.
登录
查看更多内容
影响因子：
16
作者：
Victoria Neguembor, Maria;Martin, Laura;Castells-Garcia, Alvaro;Aurelio Gomez-Garcia, Pablo;Vicario, Chiara;Carnevali, Davide;Abed, Jumana AlHaj;Granados, Alba;Sebastian-Perez, Ruben;Sottile, Francesco;Solon, Jerome;Wu, Chao-ting;Lakadamyali, Melike;Cosma, Maria Pia
通讯作者：
Cosma, Maria Pia
影响因子：
3
作者：
Rueden CT;Schindelin J;Hiner MC;DeZonia BE;Walter AE;Arena ET;Eliceiri KW
通讯作者：
Eliceiri KW
影响因子：
3.3
作者：
Grézy A;Chevillard-Briet M;Trouche D;Escaffit F
通讯作者：
Escaffit F
影响因子：
8
作者：
Jeanblanc, M.;Ragu, S.;Mann, C.
通讯作者：
Mann, C.
影响因子：
56.9
作者：
Huang, Bo;Wang, Wenqin;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei