研究一种快速准确测定微藻中中性脂的方法。湛江等鞭金藻是一种中性脂含量高且具有开发潜力的能源微藻。以湛江等鞭金藻为实验对象，首先优化尼罗红染色的条件。当二甲基亚砜体积分数为2�0％、尼罗红质量浓度为1�00μg／mL、细胞密度为1�0×106个／mL、激发波长为480 nm、检测波长为580 nm时，优化的染色时间为10 min。其次测定了背景荧光对检测的影响。结果表明，在不同细胞状态下，背景荧光强度大约是微藻内荧光强度的20％左右，可以忽略。最后比较了尼罗红荧光法和重量法。结果表明，荧光强度与中性脂含量的相关系数 R2＝0�9468，虽然两者相关性并不十分高，但作为一种快速测定微藻中中性脂的方法，尼罗红荧光法依然是研究微藻培养过程中中性脂含量变化的有效方法。%A rapid method for quantifying neutral lipids in microalgae is impending�The marine microalga Isochrysis zhangjiangensis, an ideal candidate for producing biodiesel, was taken as a model system to establish the nile red florescence method�The optimal staining condition was 2�0% dimethyl sulfoxide (V/V),1�00 μg/mL nile red,1�0×106 cell/mL cell density and staining for 10 min�The excitation and emission wavelength was 480 nm and 580 nm,respectively�Background fluorescence at different growth phase was measured and its intensity was only about 20% of the florescence intensity in cell�Compared with the fluorescence intensity in cell,the intensity of background florescence was weak and negligible�A correlation coefficient of R2=0�946 8 was obtained between the fluorescence intensity of nile red and the neutral lipid content measured by conventional gravimetric method�Although the correlation coefficient was not perfect, nile red method is useful in determining neutral lipid content during microalga cultivation.
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