Live-cell imaging was essentially performed as described (Barrales et al., 2016 (link)). In brief, cells were grown overnight in rich medium (YES) to the logarithmic phase (OD600 = 0.4–0.6). Before imaging, cells were attached to lectin (Sigma) coated glass-bottom dishes containing a microwell (MatTek). Cells were imaged using a Zeiss AxioObserver Z1 confocal spinning disk microscope with an EMM-CCD camera (Photometrics, Evolve 512) through a Zeiss Alpha Plan/Apo ×100/1.46 oil DIC M27 objective lens. Z-stacks were obtained at focus intervals of 0.4 μm. FiJi/ImageJ software was used to count the number of foci in the yeast cells.
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Live-Cell Imaging of Yeast Foci
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Corresponding Organization :
Other organizations : Max Planck Institute of Biochemistry, Ludwig-Maximilians-Universität München, University of Giessen, HUN-REN Szegedi Biológiai Kutatóközpont, Instituto de Agrobiotecnología del Litoral, National University of the Littoral, San Francisco Foundation, University of California, San Francisco, Centro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide, Junta de Andalucía, Consejo Superior de Investigaciones Científicas
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Variable analysis
independent variables
- None explicitly mentioned
- Number of foci in the yeast cells
- Growth of cells in rich medium (YES) to the logarithmic phase (OD600 = 0.4–0.6)
- Attachment of cells to lectin (Sigma) coated glass-bottom dishes containing a microwell (MatTek)
- Imaging using a Zeiss AxioObserver Z1 confocal spinning disk microscope with an EMM-CCD camera (Photometrics, Evolve 512) through a Zeiss Alpha Plan/Apo ×100/1.46 oil DIC M27 objective lens
- Acquisition of Z-stacks at focus intervals of 0.4 μm
- Use of FiJi/ImageJ software to count the number of foci in the yeast cells
- No positive or negative controls were explicitly mentioned in the provided information.
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