| Bioactivity | Nile red (Nile blue oxazone) is a lipophilic stain. Nile red has environment-sensitive fluorescence. Nile red is intensely fluorescent in a lipid-rich environment while it has minimal fluorescence in aqueous media. Nile red is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytof uorometry. Nile red stains intracellular lipid droplets red. The fluorescence wavelength is 559/635 nm[1]. Storage: protect from light. |
| Invitro | 1.1 制备储存液用 DMSO 配制 1mM 储存液。1.2 工作液的配制用预热好的无血清细胞培养基或 PBS 稀释储存液，终浓度为 200-1000 nM。注：请根据实际情况调整 Nile Red 工作液浓度，且现用现配。2.细胞染色2.1 悬浮细胞：离心收集细胞，加入 PBS 洗涤两次，每次 5 分钟。贴壁细胞：弃去培养基，加入胰蛋白酶消化细胞。离心弃去上清后，加入 PBS 洗涤两次，每次 5 分钟。2.2 加入 1 mL Nile Red 工作液，室温孵育 5-10 分钟。2.3 400 g，4℃ 离心 3-4 分钟，弃去上清。2.4 加入 PBS 洗涤细胞两次，每次 5 分钟。2.5 用 1 mL 无血清培养基或 PBS 重悬细胞后，使用荧光显微镜进行观察。 |
| In Vivo | When Nile red-stained Caenorhabditis elegans is viewed for green fluorescence, discrete lipid bodies can be observed throughout the intestine and other tissues either in clusters or evenly dispersed, depending on the animal's genotype or experimental treatment[3]. |
| Name | Nile Red |
| CAS | 7385-67-3 |
| Formula | C20H18N2O2 |
| Molar Mass | 318.37 |
| Appearance | Solid |
| Transport | Room temperature in continental US; may vary elsewhere. |
| Storage | 4°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light) |
| Reference | [1]. Greenspan P, et al. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965-73. [2]. Greenspan P, et al. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965-73. [3]. Gibrán S Alemán-Nava, et al. How to use Nile Red, a selective fluorescent stain for microalgal neutral lipids. J Microbiol Methods. 2016 Sep;128:74-79. [4]. Wilber Escorcia, et al. Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining. J Vis Exp. 2018 Mar 5;(133):57352. [5]. Elizabeth C Pino, et al. Biochemical and high throughput microscopic assessment of fat mass in Caenorhabditis elegans. J Vis Exp. 2013 Mar 30;(73):50180. |