Staining and embedding of human chromosomes for 3-d serial block-face scanning electron microscopy.

Mohammed Yusuf, Bo Chen, Teruo Hashimoto, Ana Katrina Estandarte, George Thompson, Ian Robinson

BioTechniques 2014 Dec

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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.

Citation

Mohammed Yusuf, Bo Chen, Teruo Hashimoto, Ana Katrina Estandarte, George Thompson, Ian Robinson. Staining and embedding of human chromosomes for 3-d serial block-face scanning electron microscopy. BioTechniques. 2014 Dec;57(6):302-7