Detecting Cell Cycle Stage and Progression in Fission Yeast, Schizosaccharomyces pombe

Abstract

We have previously described methods to synchronize cultures of fission yeast, Schizosaccharomyces pombe. In this chapter, we provide methods to detect cell cycle stage in cells and populations of S. pombe. These protocols used fixed samples. First, we describe sample preparation for flow cytometry of bulk DNA content. This technique allows users to monitor progression of DNA replication and detect any perturbation during the synthesis (S) phase of the cell cycle. Second, we describe methods to stain nuclei and septa of fixed yeast cells, and monitor proportions of cell cycle stages within cultures. Together, these methods provide the ability to compare cell cycle progression or delay between cultures, making use of the powerful molecular genetics tool that is S. pombe.

Keywords: Aniline blue; Cell cycle; DAPI nuclear stain; Fission yeast; Flow cytometry; S-phase; Schizosaccharomyces pombe; septa.