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.

Solid

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。

Medlife has not independently confirmed the accuracy of these methods. They are for reference only.

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 animals genotype or experimental treatment.

Medlife has not independently confirmed the accuracy of these methods. They are for reference only.

Room temperature or refrigerated transportation.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

• Description: Nile Red is a hydrophobic fluorescent for cellular lipid with red fluorescence.
  Method: For cellular lipid staining.
  1. Testing cells are inoculated in 12-well culture plates and cultured for 24 h.
  2. Stain cells with Nile Red.
  3. Use a confocal microscope (Olympus Fluoview FV1000) to visualize the lipids.
• Description: Nile Red is a hydrophobic fluorescent for cellular lipid with red fluorescence.
• Description: Nile Red is a hydrophobic fluorescent for cellular lipid with red fluorescence.
  Method: For cell staining.
  1. Cells are fixed with 4% paraformaldehyde for 1 hour at room temperature and washed 3 times with 1% BSA in PBS.
  2. Stain cells with Nile red (10 μg/mL; 4°C; overnight).
  3. Use a confocal microscope (Olympus IX81-FV1000) for image.
• Description: Nile Red is a hydrophobic fluorescent for cellular lipid with red fluorescence, it can be used in intravital imaging.
  Method: For lipid droplets staining in vivo.
  1. Inject testing mice with Nile Red (5 g) into the tail vein.
  2. Use a intravital microscopy for image. Intravital microscopy is performed with an LSM880 multi-photon-photon laser-scanning microscope (Zeiss, Oberkochen, Germany) equipped with an Olympus focus drive and motorized stage.
• Description: Nile Red is a hydrophobic fluorescent for cellular lipid with red fluorescence.
  Method: For cellular lipid staining.
  1. Fix cells with paraformaldehyde (4%; 15 min).
  2. Incubate cells with Nile Red (0.1 μg/mL; 30 min).
  3. Images are acquired with a Nicon A1 confocal microscope and processed and analysed with ImageJ software.

• [1]. Greenspan P, et al. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965-73.  [Information]

  [2]. Greenspan P, et al. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965-73.  [Information]

  [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.  [Information]

  [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.  [Information]

  [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.  [Information]

产品不同，其溶解度不同。建议根据产品选择合适的溶剂配制储备溶液；配成溶液后，建议分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C 储存时，建议在 6 个月内使用，-20°C 储存时，建议在 1 个月内使用。