基于尼罗红荧光染色的小球藻脂质快速检测方法研究

尼罗红荧光染色的脂质测定方法已经应用到一些种类的微藻当中,但在细胞壁较厚的一些绿藻中因不易染色而无法应用.选用了4株小球藻,对尼罗红荧光染色测定脂质的方法进行改进,采用20％二甲基亚砜溶液作为渗透剂,在35 ～ 40℃下对藻细胞进行前处理,加强尼罗红与胞内脂质的结合；藻细胞密度的OD540在0.8～1.1范围内,加入15 μL质量浓度为0.1 mg/mL的尼罗红丙酮溶液染色可以有效提高荧光发射强度.该方法能够较为准确地反映胞内脂质含量,可以作为自然界中广泛筛选富脂绿藻的快速检测方法.

Rapid determination of lipid in Chlorella based on Nile red fluorescence

The Nile red fluorescence method has been successfully applied to determine the lipids in certain microalgae,but it could not be applied in those with thick cell walls.The solvent of dimethyl sulfoxide(DMSO)was introduced to four strains of Chlorella samples as the penetrating agent at an elevated temperature to enhance the combination of Nile red and lipid.In the improved Nile red fluorescence staining method,the microalgae cell with OD540 of 0.8-1.1 was pretreated with 20% DMSO at 35-40 ℃ by water bath,and dyed by adding 15 μL Nile red(concentration 0.1 mg/mL).The fluorescence intensity with the improved method was enhanced.The improved Nile red fluorescence staining method could be used as a rapid detection technique for screening of naturally-occurring algal strains in a broader spectrum for high lipid production.