Fission yeast a cellular model well suited for electron microscopy investigations.
The fission yeast Schizosaccharomyces pombe has become a prominent model in molecular biology, both in yeast genetics and to investigate the molecular mechanism of the cell cycle. It has also proved to be a suitable model organism for looking at cell architecture and ultrastructure using electron microscopy (EM). Here we discuss what makes S. pombe particularly suited to EM and summarize the important discoveries regarding cell organization that have emerged from such studies. We describe the procedures and conventional methods used in EM analysis of fission yeast cells, and lay particular emphasis on cryogenic procedures, which preserve the cell structure in a near-native state, allowing elaborate three-dimensional reconstruction using electron tomography. The chapter also gives several examples of how contemporary EM approaches can be applied to provide a detailed read-out of phenotypes in this versatile cell system. A list of instruments and detailed protocols are provided together with EM-specific reagents required for sample preparation. Finally, potential new avenues of research are discussed, anticipating forthcoming topics in EM as well as new approaches to fission yeast research in the future.
Roque
Hélio
H
Cell Biology and Biophysics Program, European Molecular Biology Laboratories, Heidelberg 69117, Germany.
Antony
Claude
C
D055032:Electron Microscope Tomography / Q000295:instrumentation / Q000379:methods; D017110:Freeze Substitution / Q000379:methods; D016591:Histocytological Preparation Techniques / Q000295:instrumentation / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D012568:Schizosaccharomyces / Q000648:ultrastructure*