REQUIMTE, Department of Biological Sciences, Laboratory of Microbiology, Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal.
Folia Microbiol (Praha). 2013 Jan;58(1):81-4. doi: 10.1007/s12223-012-0186-6. Epub 2012 Jul 25.
An increasing number of researchers are using the Saccharomyces cerevisiae chronological aging model to gain insight into the post-mitotic cellular aging. Recently, an alternative approach to the traditional cellular viability assay by colony-forming unit (CFU) counts, based on the propidium iodide (PI) staining combined with flow cytometry (PI-FCM), was proposed for the assessment of yeast chronological aging. Since the chronological aging assessment shows variations particularly concerning the aging media, in this work, the influence of the most common aging media (exhausted media or water) on the assessment of chronological aging by PI staining was studied. Our results show that this methodology is highly affected by the aging media. Indeed, a correlation between CFU counts and the percentage of PI-stained cells is only achieved with the exhausted media. As such, the assessment of yeast chronological aging by PI-FCM water should not be used.
越来越多的研究人员正在使用酿酒酵母时序老化模型来深入了解有丝分裂后细胞的衰老。最近,一种替代传统细胞活力检测方法的方法,基于碘化丙啶(PI)染色结合流式细胞术(PI-FCM),用于评估酵母时序老化。由于时序老化评估特别受老化介质的影响,在这项工作中,研究了最常见的老化介质(耗尽介质或水)对 PI 染色评估时序老化的影响。我们的结果表明,这种方法受到老化介质的高度影响。实际上,只有在耗尽介质中才能实现 CFU 计数和 PI 染色细胞百分比之间的相关性。因此,不应该使用 PI-FCM 水来评估酵母的时序老化。