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首页> 外文期刊>African Journal of Biotechnology >Development of an indirect method of microalgal lipid quantification using a lysochrome dye, Nile red
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Development of an indirect method of microalgal lipid quantification using a lysochrome dye, Nile red

机译：间接溶血色素染料尼罗红微藻脂质定量方法的开发

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Earlier studies showed that the lipophilic dye, Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) can be used to measure the lipid content of microalgae by cellular?in vivo?fluorescence. It was observed that a higher amount of lipid present in lipid droplets of microalgal cells would result in higher degree of emitted fluorescent light. In this present study, the feasibility of using Nile red, a fluorescent dye specific for intracellular lipid droplets, as an indirect method of lipid quantifications was investigated. Following cellular staining of different microalgal species with nile red, the?in vivo?fluorescence of the whole cell was visualized by fluorescence microscopy (excitation: 450 to 590 nm and emission: 520 nm). Intensity of the relative?in vivofluorescence was measured using a fluorescence spectrophotometer at excitation and emission wavelengths of 485 and 590 nm, respectively. Lipid content was determined gravimetrically and the fluorescence of the extract was measured using the microemulsion method at emission and excitation wavelengths of 540 and 617 nm. The equivalent oil content of the extracted lipid was correlated to the fluorescence of pure olive oil using the microemulsion method. Cellular?in vivo?fluorescence of stained cells (ex: 485 nm and em: 590 nm), fluorescence of extracted lipid (ex: 540 nm and em: 617 nm) and gravimetrically determined lipid were linearly correlated. This suggests that Nile red can serve as a vital stain which allows a relatively rapid method of determining the lipid content of microalgal samples and is as good as the gravimetric method used for lipid determination, eliminating the requirement for the toxic solvents and time-consuming manipulations.
机译：较早的研究表明，亲脂性染料尼罗红（9-二乙基氨基-5H-苯并[α]苯恶嗪-5-酮）可用于通过细胞体内荧光检测微藻的脂质含量。观察到存在于微藻细胞的脂质液滴中的脂质的量更高将导致发出荧光的程度更高。在本研究中，研究了使用尼罗红（一种特异性针对细胞内脂质小滴的荧光染料）作为脂质定量间接方法的可行性。用尼罗红对不同的微藻种进行细胞染色后，通过荧光显微镜观察可见整个细胞的体内荧光（激发：450至590 nm，发射：520 nm）。使用荧光分光光度计分别在485和590 nm的激发和发射波长下测量相对体内荧光强度。用重量分析法测定脂质含量，并使用微乳法在540nm和617nm的发射和激发波长下测量提取物的荧光。使用微乳液法，提取的脂质的当量油含量与纯橄榄油的荧光相关。染色细胞的细胞体内荧光（例如：485 nm和em：590 nm），提取脂质的荧光（例如：540 nm和em：617 nm）与重量分析法测定的脂质呈线性相关。这表明尼罗河红可以作为重要的染色剂，它允许相对快速地确定微藻样品中脂质的含量，并且与用于脂质测定的重量分析法一样好，从而消除了对有毒溶剂和费时的操作的要求。 。

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