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| 尼罗红染色荧光定量海洋微拟球藻胞内油脂含量 |
| Fluorescent quantitation of oil content in marine Nannochloropsis oceanica by Nile Red staining |
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| DOI： |
| 中文关键词: 海洋微拟球藻 油脂含量 尼罗红 荧光强度 |
| 英文关键词:Nannochloropsis oceanica oil content Nile Red fluorescence intensity |
| 基金项目:国家自然科学基金项目（21676228）；湖南省教育厅重点项目（18A068） |
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| 中文摘要: |
| 以海洋微拟球藻Nannochloropsis oceanica为受试对象，采用尼罗红染色荧光定量胞内油脂含量。采用单因素实验考察了激发波长、发射波长、二甲基亚砜体积分数、染色时间、尼罗红质量浓度对微藻胞内油脂荧光强度的影响，在此基础上，采用响应面实验进行染色荧光定量条件优化。结果表明，优化的微拟球藻胞内油脂尼罗红染色荧光定量最佳条件为二甲基亚砜体积分数25.6%、染色时间16 min、尼罗红质量浓度0.11 μg/mL、激发波长430 nm、发射波长685 nm。在最佳条件下，尼罗红染色微拟球藻胞内油脂荧光强度与油脂含量显著正相关（R2=0999 9），说明通过尼罗红染色荧光法可快速表征微拟球藻胞内油脂含量。研究结果为后续微拟球藻资源高值化应用及高产油脂海洋微藻快速筛选和动态追踪微藻产油过程提供了依据。 |
| 英文摘要: |
| The oil content of Nannochloropsis oceanica was quantified using Nile Red staining and fluorescence method. The effects of excitation wavelength, emission wavelength, dimethyl sulfoxide (DMSO)volume fraction, Nile Red staining time and Nile Red mass concentration on fluorescence intensity of N. oceanica oil were investigated by single factor experiment. Then, the response surface methodology was used to optimize the Nile Red staining conditions as follows: excitation wavelength 430 nm, emission wavelength 685 nm,DSMO volume fraction 25.6%, Nile Red staining time 16 min, Nile Red mass concentration 0.11 μg/mL. Under the optimal conditions, the oil content of N. oceanica was positively correlated with fluorescence intensity (R2=0.999 9),which demonstrated that Nile Red staining and fluorescence quantitation could be used to rapidly and exactly detect the oil content in N. oceanica. The outcomes would be applied in high value application of N. oceanica, screening microalgal strains with higher oil content in large scale and dynamically tracking the oil-producing process of microalgae. |
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