Three-Dimensional Maps of All Chromosomes in Human Male Fibroblast Nuclei and Prometaphase Rosettes
(A) Karyotype from a female human lymphocyte (46, XX). Chromosomes were hybridized with a probe for Alu sequences (green) and counterstained with TOPRO-3 (red). Alu sequences were used as a marker for chromosomes and chromosome bands rich in genes.
(B and C) Confocal serial sections were obtained from a human G0 fibroblast nucleus (B) and a G0 lymphocyte nucleus from peripheral blood (C) after 3D FISH with the Alu probe (green) and TOPRO-3 counterstaining (red). As examples, sections made at the top, middle, and bottom of the nuclei (separated by about 1 μm) are shown from left to right. Scale bars, 5 μm.
(D) Enlarged confocal mid-section through the human G0 fibroblast nucleus. Scale bar, 5 μm.
(E) Enlargement of the boxed sector in (D). The color image in the middle reflects the merged images left (TOPRO-3 counterstaining, red) and right (Alu staining, green). Arrows indicate chromatin rich in Alu sequences expanding into the TOPRO-3-stained, Alu-poor nuclear rim. Scale bar, 2 μm.