Cargando…

A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells

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 pro...

Descripción completa

Detalles Bibliográficos
Autores principales: Martin, Laura, Vicario, Chiara, Castells-García, Álvaro, Lakadamyali, Melike, Neguembor, Maria Victoria, Cosma, Maria Pia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8488755/
https://www.ncbi.nlm.nih.gov/pubmed/34632419
http://dx.doi.org/10.1016/j.xpro.2021.100865
Descripción
Sumario: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 Neguembor et al. (2021).