Matsuoka Hideaki, Nakano Koichiro, Takatani Norimasa, Yoshida Tomonori, Igimi Shizunobu, Saito Mikako
J AOAC Int. 2014 Mar-Apr;97(2):479-83. doi: 10.5740/jaoacint.13-302.
Standard materials of a small defined number of cells with colony-forming potentiality are essential for the rational validation of food microbiological methods. An in situ flow cytometric method using viable staining with 6-carboxyfluorescein diacetate (CFDA) and tryptic soy agar (TSA) was previously proposed and its feasibility was demonstrated with five strains. In this study, this method was applied to 16 strains to support its broad applicability. The cell sorting gate was previously determined based on the CFDA stainability alone. Now the structural properties of cells designated by forward and side-scattering intensities have been introduced as the second gating criteria. Under the optimum gate condition, 100 cells have been selected and sorted on TSA. Consequently, a 95% or higher colony-forming rate has been attained for every strain. A successful application to microaerophilic Campylobacter spp. is especially of great importance because it suggests further broader applicability.
具有集落形成潜力的少量特定细胞的标准材料对于合理验证食品微生物方法至关重要。先前提出了一种使用6-羧基荧光素二乙酸酯（CFDA）活细胞染色和胰蛋白胨大豆琼脂（TSA）的原位流式细胞术方法，并用五种菌株证明了其可行性。在本研究中，该方法应用于16种菌株以支持其广泛适用性。细胞分选门先前仅基于CFDA染色性确定。现在，由前向和侧向散射强度指定的细胞结构特性已被引入作为第二个门控标准。在最佳门控条件下，已在TSA上选择并分选了100个细胞。因此，每种菌株的集落形成率均达到95%或更高。成功应用于微需氧弯曲杆菌属尤其重要，因为这表明其适用性更广泛。
