Nile Red (solution) 

Nile Red (solution) 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].
Solvent and concentration: DMSO: 1 mM

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
1. Preparation of working solution
Dilute the stored solution with preheated serum-free cell culture medium or PBS to a final concentration of 200-1000 nM.
Note: Please adjust the concentration of Nile Red working solution according to the actual situation, and prepare it as needed.
2. Cell staining
2.1 Suspension cells: Centrifuge to collect cells, add PBS for two washes, each for 5 minutes.
Adherent cells: Discard the culture medium, add trypsin to digest the cells. Centrifuge and discard the supernatant, then add PBS for two washes, each for 5 minutes.
2.2 Add 1 mL of Nile Red working solution, incubate at room temperature for 5-10 minutes.
2.3 Centrifuge at 400 g for 3-4 minutes at 4℃, discard the supernatant.
2.4 Add PBS for two washes of the cells, each for 5 minutes.
2.5 Resuspend the cells in 1 mL of serum-free medium or PBS, and observe using a fluorescence microscope.

MedChemExpress (MCE) 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 animal's genotype or experimental treatment^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

318.38

C₂₀H₁₈N₂O₂

7385-67-3

Liquid (Density: 1.2 g/cm³)

Orange to red

O=C1C2=CC=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=C1

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. 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.  [Content Brief]

  [2]. 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.  [Content Brief]

  [3]. 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.  [Content Brief]

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