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  • Title: Smart specimen preparation for freeze substitution and serial ultrathin sectioning of yeast cells.
    Author: Yamaguchi M, Okada H, Namiki Y.
    Journal: J Electron Microsc (Tokyo); 2009 Aug; 58(4):261-6. PubMed ID: 19289851.
    Abstract:
    A smart and efficient method for freeze substitution and serial sectioning of yeast cells is described. Yeast cells were placed in a single layer between two copper disks, rapidly frozen, freeze substituted and embedded in an epoxy resin. The cell layer was re-embedded by the same resin, the surface trimmed leaving 1 mum above the cell layer, and serially sectioned. The sections were collected on the two-slit grids and placed on a Formvar film mounted to cover the holes of an aluminum supporting rack. The grids were removed from the rack, stained together using a silicon tube and observed in a transmission electron microscope. The images of yeast cells observed were clear and natural, and would be useful for a detailed 3D structural analysis such as structome.
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