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Nile red is a lipophilic stain. Nile red stains intracellular lipid droplets yellow. In most polar solvents, Nile red will not fluoresce; however, when in a lipid-rich environment, it can be intensely fluorescent, with varying colors from deep red to strong yellow-gold emission. The dye is highly solvatochromic and its emission and excitation wavelength both shift depending on solvent polarity and in polar media will hardly fluoresce at all.
Nile red has applications in cell biology, where it can be used as a membrane dye which can be readily visualized using an epifluorescence microscope with excitation and emission wavelengths usually shared with red fluorescent protein. Nile red has also been used as part of a sensitive detection process for microplastics in bottled water. Additionally, nile red is a remarkable candidator in fabricating membrane for different sensors to detect environmental changes, such as taste, gas, pH, etc.
In triglycerides , Nile red has an excitation maximum of about 515 nm , and emission maximum of about 585 nm. In contrast, in phospholipids , Nile red has an excitation maximum of about 554 nm , and an emission maximum of about 638 nm.