Stockert J C, Abasolo M I, Blázquez-Castro A, Horobin R W, Revilla M, Lombardo D M
Department of Biology, Faculty of Sciences, Autonomous University of Madrid, Madrid 28049, Spain.
Biotech Histochem. 2010 Oct;85(5):277-83. doi: 10.3109/10520290903196183.
We evaluated a number of lipophilic dyes and fluorochromes, including oxazone and thiazone derivatives of oxazine and thiazine dyes, scintillator agents, a carotenoid and a metal-porphyrin complex for visualization of lipid droplets within aldehyde fixed cultured HeLa and BGC-1 cells. Observation under ultraviolet, blue or green exciting light revealed selective fluorescence of lipid droplets, particularly after treatment with aqueous solutions of Nile blue and brilliant cresyl blue oxazones, toluidine blue thiazone, or propylene glycol solutions of canthaxanthin, ethyl-BAO, and ZnTPyP. Mounting in water was required to maintain the fluorescence of lipids; the use of glycerol, Mowiol or Vectashield was not adequate. The effect of dye structure on staining intensity was assessed with the aid of numerical structure parameters modeling lipophilicity (HI and log P), overall size (MW) and the size of the conjugated system (conjugated bond number; CBN). The best stains for lipid droplets were relatively lipophilic (HI > 4.0, log P > 5.0), of small size overall (MW < 370), with small conjugated systems (CBN < 24), and not significantly amphiphilic. The two hydrophobic-hydrophilic parameters (the classic log P and the hydrophobic index, HI; values calculated by molecular modeling software) were highly correlated; however, HI was a more suitable hydrophobicity index for the dyes studied here.
我们评估了多种亲脂性染料和荧光染料，包括恶嗪和噻嗪染料的恶唑酮和噻唑酮衍生物、闪烁剂、一种类胡萝卜素和一种金属卟啉配合物，用于在醛固定的培养HeLa细胞和BGC-1细胞中可视化脂滴。在紫外光、蓝光或绿光激发光下观察发现脂滴有选择性荧光，特别是在用尼罗蓝和亮甲酚蓝恶唑酮、甲苯胺蓝噻唑酮的水溶液，或角黄素、乙基-BAO和ZnTPyP的丙二醇溶液处理后。需要用水封片以维持脂质的荧光；使用甘油、Mowiol或Vectashield是不够的。借助模拟亲脂性（疏水指数HI和log P）、整体大小（分子量MW）和共轭体系大小（共轭键数；CBN）的数值结构参数评估染料结构对染色强度的影响。用于脂滴的最佳染料相对亲脂（HI > 4.0，log P > 5.0），整体尺寸较小（MW < 370），共轭体系较小（CBN < 24），且无明显两亲性。两个疏水-亲水参数（经典的log P和疏水指数HI；由分子模拟软件计算的值）高度相关；然而，HI是此处研究的染料更合适的疏水性指数。
