Quantitative DNA Methylation Imaging of Cells and Tissues

Abstract

The invention is a method using immunofluorescence, high-resolution imaging, and 3D image analysis to quantify spatial distribution of methylcytosine (MeC) in global DNA in individual cell nuclei, for the characterization of cells and tissues based on their nuclear MeC distribution patterns. The invented method is intended for basic research, clinical diagnostics and prognostics, pharmacology and toxicology, and molecular and cell-based therapy. We describe an image-cytometrical tool performing rapid cell-by-cell assay to analyze the impact of environmental conditions on global DNA methylation, including alterations of methylcytosine load and the spatial reorganization of methylated DNA in cell nuclei. The assay can be combined with molecular approaches for profiling of cells and their genomes. In contrast to molecular methods, it provides information regarding the location of MeC in cell nuclei. The resulting assay can analyze thousands of cells in parallel in a cost-effective manner—currently not possible with molecular methods.

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