Garcia M T, Sanz R, Galceran M T, Puignou L
Departament de Química Analítica, Universitat de Barcelona, Martí i Franquès, 1-11, 08028 Barcelona, Spain.
Biotechnol Prog. 2006 May-Jun;22(3):847-52. doi: 10.1021/bp050421q.
The quality of wine greatly depends on the features of the yeast used in its production, and yeast cell viability is one of the most important quality control issues to consider in this regard. In the first steps of winemaking, the use of a low-cost and simple methodology for monitoring the cell viability of yeast inoculates is of paramount importance. Gravitational field-flow fractionation is a useful technique for the determination of cell viability because it provides gentle experimental conditions, although the proper use of fluorophore probes as biomass indicators is required. In this paper the use of different fluorescent probes such as carboxyfluorescein diacetate (cFDA), calcein-AM, and SYTO-13 were considered as viability biomarkers. Calceina-AM allowed the establishment of a direct GrFFF method to determine cell viability, with a limit of detection of 5.0 x 10(4) viable cell/mL. SYTO-13 could be used as biomass indicator with a limit of detection of 3.5 x 10(4) total cells/mL. The suitability of the procedure was tested with three commercial yeast samples, and the results were compared with those obtained using standard techniques.
葡萄酒的品质很大程度上取决于其生产过程中所使用酵母的特性，而酵母细胞活力是这方面需要考虑的最重要的质量控制问题之一。在酿酒的最初阶段，采用一种低成本且简单的方法来监测酵母接种物的细胞活力至关重要。重力场流分离法是一种用于测定细胞活力的有用技术，因为它能提供温和的实验条件，不过需要恰当地使用荧光团探针作为生物量指标。在本文中，不同的荧光探针如羧基荧光素二乙酸酯（cFDA）、钙黄绿素-AM和SYTO-13被视为活力生物标志物。钙黄绿素-AM使得建立一种直接的重力场流分离法来测定细胞活力成为可能，其检测限为5.0×10⁴个活细胞/毫升。SYTO-13可用作生物量指标，检测限为3.5×10⁴个总细胞/毫升。用三个商业酵母样品对该方法的适用性进行了测试，并将结果与使用标准技术获得的结果进行了比较。
