Eclipse e200 optical

The total lifespan was defined as the length of life of a single mother cell expressed in units of time. The total lifespan was calculated as the sum of reproductive and post-reproductive lifespans. The reproductive lifespan was defined as the length of time between the first and the last budding and the post-reproductive lifespan as the length of time from the last budding until cell death. The lifespan of the Saccharomyces cerevisiae yeast was determined as previously described by [38 (link)] with small modification [37 (link)]. Ten microliter aliquots of an overnight grown culture of yeast were collected and transferred on YPD, YPGal, or SD-Ura plates with solid medium containing Phloxine B (10 μg/mL). Phloxine B was used to stain Saccharomyces cerevisiae dead cells. Dead yeast cells lost membrane integrity and Phloxine B entered cell space giving pink/red coloration of cytosol. In each experiment, 45 single cells were analyzed. During manipulation, the plates were kept at 28 °C for 15 h and at 4 °C during the night. The results represent measurements for at least 90 cells analyzed in two independent experiments. The analysis was performed by micromanipulation using the Nikon Eclipse E200 optical microscope with an attached micromanipulator.

The total lifespan is the length of life of a single mother yeast cell expressed in units of time and was calculated as the sum of reproductive (time between the first and the last budding) and post-reproductive lifespans (time from the last budding until cell death). The total lifespan of the S. cerevisiae yeast was determined as previously described in [18 (link)] with small modifications from [36 (link)]. Ten-microliter aliquots of a fresh exponential culture of yeast were collected and transferred on YPD plates with solid medium containing Phloxine B (10 μg/mL). In each experiment, 45 single cells were analyzed. During manipulation, the plates were kept at 28 °C for 15 h and at 4 °C during the night. The results represent measurements for at least 90 cells analyzed in at least two independent experiments. The analysis was performed by micromanipulation using a Nikon Eclipse E200 optical microscope with an attached micromanipulator.

The budding lifespan of individual mother yeast cells was defined as the number of mitotic cycles (buddings) during the cell’s life. After overnight growth, cells were arrayed on a YPD plate without (control) or with 200 μM and 300 μM curcumin using a micromanipulator. The budding lifespan was determined microscopically by a routine procedure with the use of a micromanipulator as described previously (Molon et al. 2018 (link)). The number of buds formed by each mother cell signifies its reproductive potential (budding lifespan). In each experiment, forty-five single cells were analyzed. The results represent measurements for at least 90 cells analyzed in two independent experiments. The analysis was performed by micromanipulation using the Nikon Eclipse E200 optical microscope with an attached micromanipulator.