Nile Red

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 543/598 nm^[1]. In Vitro: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 storage solution and working solution
1.1 Preparation of storage solution
Prepare 1 mM storage solution using DMSO.
1.2 Preparation of working solution
Dilute the storage solution with pre-heated 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.
2.3 400 g, 4℃ centrifuge for 3-4 minutes, 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.
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].