Ultrastructural Visualization of 3D Chromatin Folding Using Serial Block-Face Scanning Electron Microscopy and In Situ Hybridization (3D-EMISH)
| Autor: | Grzegorz M. Wilczynski, Andrzej A. Szczepankiewicz, Michal Kadlof, Agnieszka Walczak, Artur Wolny, Blazej Ruszczycki, Parteka Z, Katarzyna Karolina Pels, Jesse Aaron, Małgorzata Alicja Śliwińska, Grzegorz Bokota, Sebastian Arabasz, Magdalena Kiss-Arabasz, Pawel Trzaskoma, Adriana Magalska, Dariusz Plewczynski, Yijun Ruan, Katarzyna Krawczyk, Byoungkoo Lee |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Serial block-face scanning electron microscopy
0303 health sciences Chemistry Resolution (electron density) Folding (DSP implementation) Genome law.invention Chromatin 03 medical and health sciences 0302 clinical medicine law Ultrastructure Biophysics Human genome Electron microscope 030217 neurology & neurosurgery 030304 developmental biology |
| DOI: | 10.1101/2020.02.05.935106 |
| Popis: | The human genome is extensively folded into 3-dimensional organization, yet the detailed 3D chromatin folding structures have not been fully visualized due to the lack of robust and ultra- resolution imaging capability. Here, we report the development of a novel electron microscopy method that combines serial block-face scanning electron microscopy with in situ hybridization (3D-EMISH) to visualize 3D chromatin folding at targeted genomic regions with ultra-resolution (5×5×30 nm in xyz dimensions, respectively). We applied 3D-EMISH to human lymphoblastoid cells at a 1.7 Mb segment of the genome and visualized a large number of distinctive 3D chromatin folding structures in high ultra-resolution. We further quantitatively characterized the reconstituted chromatin folding structures by identifying sub-domains, and uncovered a high level of heterogeneity in chromatin folding ultrastructures, suggestive of extensive dynamic fluidity in 3D chromatin states. |
| Databáze: | OpenAIRE |
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