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 559/635 nm^[1]. In Vitro:1.1 Preparation of stock solution
Use DMSO to prepare 1 mM stock solution.
1.2 Preparation of working solution
Use preheated serum-free cell culture medium or PBS to dilute the stock solution 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 before use.
2. Cell staining
2.1 Suspended cells: Collect cells by centrifugation and wash twice with PBS for 5 minutes each time.
Adherent cells: Discard the culture medium and add trypsin to digest the cells. After centrifugation and discarding the supernatant, wash twice with PBS for 5 minutes each time.
2.2 Add 1 mL Nile Red working solution and incubate at room temperature for 5-10 minutes.
2.3 Centrifuge at 400 g, 4℃ for 3-4 minutes and discard the supernatant.
2.4 Wash cells twice with PBS for 5 minutes each time.
2.5 Resuspend the cells in 1 mL serum-free medium or PBS and observe under 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].