摘要
为了建立快速检测裂殖壶菌胞内油脂的方法,探讨了尼罗红荧光染色法检测裂殖壶菌油脂含量的检测条件。通过考察裂殖壶菌重悬液中添加二甲基亚砜体积分数、尼罗红染液用量、染色温度、染色时间及细胞密度对荧光强度的影响,确立最优检测条件,进一步考察细胞油脂含量与荧光强度的关系,从而建立尼罗红荧光检测法的定量关系。结果表明,最优检测条件为：每毫升裂殖壶菌重悬液中二甲基亚砜体积分数和尼罗红染液用量分别为25%和15μL,染色温度50℃,染色时间15 min。在最优检测条件下,细胞密度不超过0.218×10^8个/m L范围内,菌液油脂含量（X）与荧光强度（Y）呈现良好的线性关系,其线性关系式为Y=798.55X-3.44,相关系数R2为0.996 4。因此,采用尼罗红荧光染色法可以快速检测裂殖壶菌的油脂含量。
In order to establish a rapid method for determining the oil in Schizochytrium sp. cell, the determination conditions of oil content in Schizochytrium sp. by Nile Red fluorescence dyeing method were studied. Through investigating the effects of volume fraction of dimethyl sulfoxide and dosage of Nile Red dye solution in Schizochytrium sp. heavy suspension, dyeing temperature, dyeing time and cell density on the fluorescent intensity, the optimal determination conditions were obtained, and the correlation between the oil content in cell and fluorescent intensity was further studied so as to establish the quantitative relation of Nile Red fluorescence dyeing method. The results showed that the optimal determination conditions were obtained as follows： volume fraction of dimethyl sulfoxide and dosage of Nile Red dye solution in per milliliter of Schizochytrium sp. heavy suspension 25% and 15 μL respectively, dyeing temperature 50℃, dyeing time 15 min. Under the optimal determination conditions, when the cell density was within 0. 218 × 10^8 cells per milliliter,there was a good linear relationship between fluorescence intensity （Y） and oil content （X） of the bacteria, and the regression equation was Y = 798.55X- 3.44 （R^2 = 0.996 4）. In conclusion, the oil content in Schizochytrium sp. could be rapidly determined by Nile Red fluorescence dyeing method.
出处
《中国油脂》
CAS
CSCD
北大核心
2016年第1期51-55,共5页
China Oils and Fats
基金
科技部"十二五"农业领域科技计划项目子课题(2011BAD14B03-06)
广东省国际合作项目(2011B050400027)
广东海洋大学创新强校工程项目(GDOU2014050203)
广东海洋大学人才引进启动项目(E11096)
关键词
裂殖壶菌
油脂
尼罗红荧光染色
Schizochytrium sp.
oil
Nile Red fluorescence dyeing
