USE OF NILE RED AS A RAPID MEASURE OF LIPID-CONTENT IN CILIATES
USE OF NILE RED AS A RAPID MEASURE OF LIPID-CONTENT IN CILIATES
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DOI：
10.1016/s0932-4739(11)80129-x
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发表时间：
1990-06-29
影响因子：
2.9
通讯作者：
ALLEN, RD
中科院分区：
文献类型：
--
作者：
COLE, TA;FOK, AK;ALLEN, RD
我们报告使用尼罗河红作为一种快速和廉价的方法来估计细胞脂质草履虫和四膜虫的三个物种的直接应用的染料活细胞或固定细胞，而无需提取和纯化。使用荧光显微镜获得不同培养年龄的细胞的脂质含量的相对变化的定性估计，而半定量测定通过测量总荧光从发射光谱（激发，535 nm）的固定细胞处理过量尼罗红。在这些选定的光谱条件下，使用荧光强度近似中性（激发，488 nm;发射，540 nm）和极性（激发，535 nm;发射，680 nm）脂质的相对量，以避免彼此的任何溢出。随着培养年龄的变化模式，在四膜虫和草履虫的总，中性和极性脂质的总，中性和极性的使用尼罗红同意与出版的重量数据，这些纤毛虫。
We report the use of nile red as a rapid and inexpensive method to estimate cellular lipids in three species of Paramecium and in Tetrahymena by the direct application of the dye to living or fixed cells without extraction and purification. Qualitative estimates of the relative changes in the lipid content of cells of varying culture ages were obtained using fluorescence microscopy, while semiquantitative determinations were obtained by measuring the total fluorescence from the emission spectrum (excitation, 535 nm) of fixed cells treated with excess nile red. The relative amounts of neutral (excitation, 488 nm; emission, 540 nm) and polar (excitation, 535 nm; emission, 680 nm) lipids were approximated using fluorescence intensity at these selected spectral conditions to avoid any spill over from each other. The pattern of change with culture age in total lipids in Tetrahymena and in total, neutral and polar lipids in Paramecium obtained using nile red agreed well with published gravimetric data for these ciliates.