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. 1999 Oct 4;147(1):13–24. doi: 10.1083/jcb.147.1.13

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© 1999 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Preservation of the spatial distribution of nascent RNA, DNA, and acetylated histone H4 in human primary fibroblasts during the FISH procedure. Optical sections were obtained before (A, C, and E) and after (B, D, and F) carrying out the FISH protocol. A and B, Optical sections of nuclei in which transcription sites were immunofluorescently labeled. The distribution of nascent RNA did not change significantly during the FISH procedure. C and D, Optical sections of nuclei labeled with the fluorescent DNA stain Sytox green. The pattern of DNA staining did not change significantly after in situ hybridization. E and F, Optical sections of nuclei in which acetylated histone H4 was fluorescently labeled. The distribution did not change significantly during the chromosome painting procedure. The diffuse, low intensity labeling observed before FISH was diminished after the procedure, resulting in a somewhat more pronounced granular labeling. Images shown have been subjected to 3-D image restoration. Bar, 1.75 μm.