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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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 |
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author | Martin, Laura Vicario, Chiara Castells-García, Álvaro Lakadamyali, Melike Neguembor, Maria Victoria Cosma, Maria Pia |
author_facet | Martin, Laura Vicario, Chiara Castells-García, Álvaro Lakadamyali, Melike Neguembor, Maria Victoria Cosma, Maria Pia |
author_sort | Martin, Laura |
collection | PubMed |
description | 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). |
format | Online Article Text |
id | pubmed-8488755 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-84887552021-10-08 A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells Martin, Laura Vicario, Chiara Castells-García, Álvaro Lakadamyali, Melike Neguembor, Maria Victoria Cosma, Maria Pia STAR Protoc Protocol 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). Elsevier 2021-09-30 /pmc/articles/PMC8488755/ /pubmed/34632419 http://dx.doi.org/10.1016/j.xpro.2021.100865 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Martin, Laura Vicario, Chiara Castells-García, Álvaro Lakadamyali, Melike Neguembor, Maria Victoria Cosma, Maria Pia A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title | A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title_full | A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title_fullStr | A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title_full_unstemmed | A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title_short | A protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
title_sort | protocol to quantify chromatin compaction with confocal and super-resolution microscopy in cultured cells |
topic | Protocol |
url | 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 |
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