Smart specimen preparation for freeze substitution and serial ultrathin sectioning of yeast cells.
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.
Yamaguchi
Masashi
M
Medical Mycology Research Center, Chiba University, Chuo-ku, Chiba, Japan. yama@faculty.chiba-u.jp
Okada
Hitoshi
H
Namiki
Yuichi
Y
D004853:Epoxy Resins; D017110:Freeze Substitution; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D008867:Microtomy / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure*; D016610:Tissue Embedding
D004853:Epoxy Resins