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
_version_ 1784578226312445952
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
work_keys_str_mv AT martinlaura aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT vicariochiara aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT castellsgarciaalvaro aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT lakadamyalimelike aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT neguembormariavictoria aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT cosmamariapia aprotocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT martinlaura protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT vicariochiara protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT castellsgarciaalvaro protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT lakadamyalimelike protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT neguembormariavictoria protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells
AT cosmamariapia protocoltoquantifychromatincompactionwithconfocalandsuperresolutionmicroscopyinculturedcells