尼罗红-荧光光谱法能够快速地对微藻细胞中的油脂含量进行检测,但是该方法对所测微藻物种具有一定依赖性,在分析不同微藻细胞油脂含量前需要对测试条件进行优化。优化后的亚心形扁藻油脂含量分析条件为:藻液800×g离心5min去除培养基,用体积分数4%的二甲基亚砜(DMSO)水溶液重悬至OD750=0.3,尼罗红质量浓度0.8μg/mL,避光40℃水浴恒温10min,激发波长520nm。发现亚心形扁藻在培养的对数生长后期油脂开始积累,稳定期后期油脂含量趋于恒定,优化后的尼罗红-荧光光谱法能够准确地检测亚心形扁藻生长过程中的油脂含量变化。
Nile red fluorescence spectrometry can quickly detect the lipid content in microalgal cells, but the method has certain dependence on microalgal species,so the detection conditions need optimizing for each speey before lipid content determination. For Platymonas subcordiformis, the analysis conditions of Nile red fluoresence spectrometry were optimized as follows : centrifugation at 800 × g for 5 min to remove culture medium,suspending the cells with 4% dimethylsulfoxide to OD750 0. 3,adding Nile red with final concentration 0.8 μg/mL, incubating the staining system at 40 ℃ for 10 min. Then the sample was measured by fluorescence spectrometer excited at 520 nm. Lipids in Platymonas subcordiformis began to accumulate at late exponential phase and reached a stable value at late stabilization period, and the optimized Nile red fluoresence spectrometry could accurately detect the relative lipid content in Platymonas subcordiformis at different growth periods.