Analysis of Cell Cycle Progression in Saccharomyces cerevisiae Using the Budding Index and Tubulin Staining.
The budding index and the morphology of the spindle and the nucleus are excellent markers for the analysis of the progression through the different stages of the cell cycle in Saccharomyces cerevisiae. Here, we describe a protocol to evaluate the budding index in this model organism using phase contrast microscopy. We also describe an indirect immunofluorescence method designed for the visualization of microtubules and the nucleus in S. cerevisiae. Finally, we explain how both methodologies can be used in order to analyze cell cycle progression in the budding yeast.
Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Consejo Superior de Investigaciones Científicas (CSIC), Avda. Américo Vespucio, s/n, P.C.T. Cartuja 93, 41092, Sevilla, Spain.
Marta
M
Muñoz-Barrera
Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Consejo Superior de Investigaciones Científicas (CSIC), Avda. Américo Vespucio, s/n, P.C.T. Cartuja 93, 41092, Sevilla, Spain. fernando.monje@cabimer.es.
Fernando
F
Monje-Casas
D002453:Cell Cycle; D005455:Fluorescent Antibody Technique / Q000379:methods*; D008858:Microscopy, Phase-Contrast / Q000379:methods*; D008870:Microtubules / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis*; D008941:Spindle Apparatus / Q000648:ultrastructure; D014404:Tubulin / Q000032:analysis*
D029701:Saccharomyces cerevisiae Proteins; D014404:Tubulin
Budding index; Cell cycle; Immunofluorescence; Microtubules; Mitotic exit; Nuclear morphology; Spindle