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

ABSTRACT: 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).

SUBMITTER: Martin L

PROVIDER: S-EPMC8488755 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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

Martin Laura L Vicario Chiara C Castells-García Álvaro Á Lakadamyali Melike M Neguembor Maria Victoria MV Cosma Maria Pia MP

STAR protocols 20210930 4

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 approach ...[more]

PMID: 34632419

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