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为探讨流式细胞术对单增李斯特菌和酿酒酵母的活菌与热灭活菌的检出效果,本文采用荧光染色试剂SYTO-9和碘化乙锭(PI)对单增李斯特菌和酿酒酵母的活菌与热灭活菌的细胞悬液进行染色,采用流式细胞仪同时测量红色荧光与绿色荧光从而得出细胞悬液中的细菌和酵母的含量。结果表明经核酸荧光染料染色后,再结合流式细胞术对细菌与酵母菌进行检测,步骤简单、耗时短。该法不仅简化了测量步骤且分辨率高,对单增李斯特菌和酿酒酵母均具有良好的检出结果,能分辨同一体系中同一菌种的活细胞与热灭活细胞和同一体系中的细菌与酵母活细胞;该法检出限低,将单增李斯特菌稀释后,最低检出限可达1.2×104 cells/mL,将酿酒酵母稀释后,最低检出限可达6×103 cells/mL,因此能大大缩短增菌时间或者避免繁复的增菌步骤。
In order to investigate the detection effect of viable cells and heat-inactivated bacteria by Listeria monocytogenes and Saccharomyces cerevisiae by flow cytometry, the fluorescence staining reagent SYTO-9 and ethidium iodide (PI) were used to detect Listeria monocytogenes And Saccharomyces cerevisiae viable cells and heat-inactivated cell suspension staining, using flow cytometry simultaneous measurement of red fluorescence and green fluorescence to arrive at the cell suspension of bacteria and yeast content. The results show that after staining with nucleic acid fluorescent dye, combined with flow cytometry to detect bacteria and yeast, the steps are simple and time-consuming. The method not only simplifies the measurement steps and high resolution, good detection of Listeria monocytogenes and Saccharomyces cerevisiae, can distinguish between the same species of living cells and heat-inactivated cells and the same system in the same system Bacteria and yeast living cells; detection limit is low, the limit of detection of Listeria monocytogenes after dilution of up to 1.2 × 104 cells / mL, the minimum detectable limit of Saccharomyces cerevisiae up to 6 × 103 cells / mL, so can greatly shorten the time of enrichment or to avoid complicated steps of enrichment.