Simultaneously defining cell phenotypes, cell cycle, and chromatin modifications at single-cell resolution

Allison B. Chambliss, Pei Hsun Wu, Wei Chiang Chen, Sean X. Sun, Denis Wirtz

Research output: Contribution to journalArticle

Abstract

Heterogeneity of cellular phenotypes in asynchronous cell populations placed in the same biochemical and biophysical environment may depend on cell cycle and chromatin modifications; however, no current method can measure these properties at single-cell resolution simultaneously and in situ. Here, we develop, test, and validate a new microscopy assay that rapidly quantifies global acetylation on histone H3 and measures a wide range of cell and nuclear properties, including cell and nuclear morphology descriptors, cell-cycle phase, and F-actin content of thousands of cells simultaneously, without cell detachment from their substrate, at single-cell resolution. These measurements show that isogenic, isotypic cells of identical DNA content and the same cell-cycle phase can still display large variations in H3 acetylation and that these variations predict specific phenotypic variations, in particular, nuclear size and actin cytoskeleton content, but not cell shape. The dependence of cell and nuclear properties on cell-cycle phase is assessed without artifact-prone cell synchronization. To further demonstrate its versatility, this assay is used to quantify the complex interplay among cell cycle, epigenetic modifications, and phenotypic variations following pharmacological treatments affecting DNA integrity, cell cycle, and inhibiting chromatin-modifying enzymes.

Original languageEnglish (US)
Pages (from-to)2667-2676
Number of pages10
JournalFASEB Journal
Volume27
Issue number7
DOIs
StatePublished - Jul 2013

Fingerprint

Chromatin
Cell Cycle
Cells
Phenotype
Nuclear properties
Acetylation
Actins
Assays
DNA
Histones
Microscopic examination
Synchronization
Cell Shape
Actin Cytoskeleton
Epigenomics
Artifacts
Microscopy
Substrates
Enzymes
Pharmacology

Keywords

  • Epigenetics
  • High-throughput
  • Microscopy

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Simultaneously defining cell phenotypes, cell cycle, and chromatin modifications at single-cell resolution. / Chambliss, Allison B.; Wu, Pei Hsun; Chen, Wei Chiang; Sun, Sean X.; Wirtz, Denis.

In: FASEB Journal, Vol. 27, No. 7, 07.2013, p. 2667-2676.

Research output: Contribution to journalArticle

Chambliss, Allison B. ; Wu, Pei Hsun ; Chen, Wei Chiang ; Sun, Sean X. ; Wirtz, Denis. / Simultaneously defining cell phenotypes, cell cycle, and chromatin modifications at single-cell resolution. In: FASEB Journal. 2013 ; Vol. 27, No. 7. pp. 2667-2676.
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