Chromatin forms nanoscale three-dimensional packing domains with fractal-like scaling behavior in vitro
Li, Y.; Agrawal, V.; Virk, R. K. A.; Roth, E. W.; Eshein, A.; Frederick, J.; Huang, K.; Almassalha, L.; Bleher, R.; Carignano, M. A.; Szleifer, I. G.; Dravid, V. P.; Backman, V.
Show abstract
Chromatin organization over a wide range of length scales plays a critical role in the regulation of transcription and deciphering the interplay of these processes requires high-resolution, three-dimensional, quantitative imaging of chromatin structure in vitro. Herein, we introduce ChromSTEM, a method that utilizes high angle annular dark-field imaging and tomography in scanning transmission electron microscopy combined with DNA-specific staining for electron microscopy. We utilized ChromSTEM to quantify chromatin structure in cultured cells and the statistical packing behavior of the chromatin polymer. Using chromatin mass and density analysis, we observed that chromatin forms spatially well-defined higher-order domains which are around 100 nm in radius, with a radially decreasing mass-density from the center to the periphery. Although the morphological properties of the domains vary within the same cell line, they seem to exhibit greater heterogeneity across cell lines, underlying a potential role of statistical chromatin packing in regulating cell-type-specific gene expression.
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