Journal article
Real-time visualization of chromatin modification in isolated nuclei
Journal of cell science, v 130(17), pp 2926-2940
01 Sep 2017
PMID: 28743737
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Chromatin modification is traditionally assessed in biochemical assays that provide average measurements of static events given that the analysis requires components from many cells. Microscopy can visualize single cells, but the cell body and organelles can hamper staining and visualization of the nucleus. Normally, chromatin is visualized by immunostaining a fixed sample or by expressing exogenous fluorescently tagged proteins in a live cell. Alternative microscopy tools to observe changes of endogenous chromatin in real-time are needed. Here, we isolated transcriptionally competent nuclei from cells and used antibody staining without fixation to visualize changes in endogenous chromatin. This method allows the real-time addition of drugs and fluorescent probes to one or more nuclei while under microscopy observation. A high-resolution map of 11 endogenous nuclear markers of the histone code, transcription machinery and architecture was obtained in transcriptionally active nuclei by performing confocal and structured illumination microscopy. We detected changes in chromatin modification and localization at the single-nucleus level after inhibition of histone deacetylation. Applications in the study of RNA transcription, viral protein function and nuclear architecture are presented. This article has an associated First Person interview with the first author of the paper.
Metrics
Details
- Title
- Real-time visualization of chromatin modification in isolated nuclei
- Creators
- Luca Sardo - University of the SciencesAngel Lin - University of the SciencesSvetlana Khakhina - University of the SciencesLucas Beckman - University of the SciencesLuis Ricon - University of the SciencesWeam Elbezanti - University of the SciencesTara Jaison - University of the SciencesHarshad Vishwasrao - National Institute of Biomedical Imaging and BioengineeringHari Shroff - National Institute of Biomedical Imaging and BioengineeringChristopher Janetopoulos - University of the SciencesZachary A Klase - University of the Sciences
- Publication Details
- Journal of cell science, v 130(17), pp 2926-2940
- Publisher
- Company of Biologists
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Pharmacology and Physiology
- Web of Science ID
- WOS:000408902000019
- Scopus ID
- 2-s2.0-85028615652
- Other Identifier
- 991021902600104721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Cell Biology