RRC ID 47026
Author Yamaguchi M, Okada H, Namiki Y.
Title Smart specimen preparation for freeze substitution and serial ultrathin sectioning of yeast cells.
Journal J Electron Microsc (Tokyo)
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.
Volume 58(4)
Pages 261-6
Published 2009-8
DOI 10.1093/jmicro/dfp013
PII dfp013
PMID 19289851
MeSH Epoxy Resins Freeze Substitution* Microscopy, Electron, Transmission / methods* Microtomy / methods* Saccharomyces cerevisiae / ultrastructure* Tissue Embedding*
Times Cited 23
Pathogenic microorganisms NA