Nuclear cytometry and chromatin organization

Smith, Paul J., Darzynkiewicz, Zbigniew and Errington, Rachel J. ORCID: https://orcid.org/0000-0002-8016-4376 2018. Nuclear cytometry and chromatin organization. Cytometry Part A 93 (8) , pp. 771-784. 10.1002/cyto.a.23521

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Abstract

The nuclear‐targeting chemical probe, for the detection and quantification of DNA within cells, has been a mainstay of cytometry—from the colorimetric Feulgen stain to smart fluorescent agents with tuned functionality. The level of nuclear structure and function at which the probe aims to readout, or indeed at which a DNA‐targeted drug acts, is shadowed by a wide range of detection modalities and analytical methods. These methods are invariably limited in terms of the resolution attainable versus the volume occupied by targeted chromatin structures. The scalar challenge arises from the need to understand the extent and different levels of compaction of genomic DNA and how such structures can be re‐modeled, reported, or even perturbed by both probes and drugs. Nuclear cytometry can report on the complex levels of chromatin order, disorder, disassembly, and even active disruption by probes and drugs. Nuclear probes can report defining features of clinical and therapeutic interest as in NETosis and other cell death processes. New cytometric approaches continue to bridge the scalar challenges of analyzing chromatin organization. Advances in super‐resolution microscopy address the resolution and depth of analysis issues in cellular systems. Typical of recent insights into chromatin organization enabled by exploiting a DNA interacting probe is ChromEM tomography (ChromEMT). ChromEMT uses the unique properties of the anthraquinone‐based cytometric dye DRAQ5™ to reveal that local and global 3D chromatin structures effect differences in compaction. The focus of this review is nuclear and chromatin cytometry, with linked reference to DNA targeting probes and drugs as exemplified by the anthracenediones.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Publisher:	Wiley
ISSN:	1552-4922
Date of First Compliant Deposit:	12 October 2018
Date of Acceptance:	24 August 2018
Last Modified:	26 Nov 2024 17:00
URI:	https://orca.cardiff.ac.uk/id/eprint/115822

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