Yusuf, M;
Chen, B;
Hashimoto, T;
Estandarte, AK;
Thompson, G;
Robinson, I;
(2014)
Staining and Embedding of Human Chromosomes for 3-D Serial Block-Face Scanning Electron Microscopy.
Biotechniques
, 57
(6)
pp. 302-307.
10.2144/000114236.
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Abstract
The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial block-face scanning electron microscopy (SBFSEM). Polyamine chromosomes are first separated using a simple filtration method and then stained with heavy metal. We show that the DNA-specific platinum blue provides higher contrast than osmium tetroxide. A two-step procedure for embedding chromosomes in resin is then used to concentrate the chromosome samples. After stacking the SBFSEM images, a familiar X-shaped chromosome was observed in 3-D.
| Type: | Article |
|---|---|
| Title: | Staining and Embedding of Human Chromosomes for 3-D Serial Block-Face Scanning Electron Microscopy |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.2144/000114236 |
| Publisher version: | http://dx.doi.org/10.2144/000114236 |
| Language: | English |
| Additional information: | This is an open access article distributed under the terms of the Creative Commons CC BY license, https://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, distribution, reproduction in any medium, provided the original work is properly cited. |
| Keywords: | Chromosome, heavy metal staining, imaging, platinum blue, serial block-face electron microscopy, three-dimensional, Cell Line, Chromosomes, Human, Coloring Agents, Histocytological Preparation Techniques, Humans, Male, Microscopy, Electron, Scanning, Mitosis, Platinum |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Chemical Engineering UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > London Centre for Nanotechnology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1465676 |
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