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Ultra-Structural Imaging Provides 3D Organization of 46 Chromosomes of a Human Lymphocyte Prophase NucleusSajid, Atiqa; email: firstname.lastname@example.org; Lalani, El-Nasir; email: email@example.com; Chen, Bo; orcid: 0000-0001-7515-3042; email: firstname.lastname@example.org; Hashimoto, Teruo; email: T.Hashimoto@Manchester.ac.uk; Griffin, Darren K.; orcid: 0000-0001-7595-3226; email: D.K.Griffin@kent.ac.uk; Bhartiya, Archana; email: email@example.com; Thompson, George; email: firstname.lastname@example.org; Robinson, Ian K.; orcid: 0000-0003-4897-5221; email: email@example.com; Yusuf, Mohammed; email: firstname.lastname@example.org (MDPI, 2021-06-01)Three dimensional (3D) ultra-structural imaging is an important tool for unraveling the organizational structure of individual chromosomes at various stages of the cell cycle. Performing hitherto uninvestigated ultra-structural analysis of the human genome at prophase, we used serial block-face scanning electron microscopy (SBFSEM) to understand chromosomal architectural organization within 3D nuclear space. Acquired images allowed us to segment, reconstruct, and extract quantitative 3D structural information about the prophase nucleus and the preserved, intact individual chromosomes within it. Our data demonstrate that each chromosome can be identified with its homolog and classified into respective cytogenetic groups. Thereby, we present the first 3D karyotype built from the compact axial structure seen on the core of all prophase chromosomes. The chromosomes display parallel-aligned sister chromatids with familiar chromosome morphologies with no crossovers. Furthermore, the spatial positions of all 46 chromosomes revealed a pattern showing a gene density-based correlation and a neighborhood map of individual chromosomes based on their relative spatial positioning. A comprehensive picture of 3D chromosomal organization at the nanometer level in a single human lymphocyte cell is presented.