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

    Research output: Contribution to journalArticlepeer-review

    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.

    Original languageEnglish
    Pages (from-to)302-307
    Number of pages6
    JournalBioTechniques
    Volume57
    Issue number6
    DOIs
    Publication statusPublished - 2014

    Keywords

    • Chromosome
    • Heavy metal staining
    • Imaging
    • Platinum blue
    • Serial block-face electron microscopy
    • Three-dimensional

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