Molecular Weight: 318.37
Storage: RT, L
[C=Cold D=Desiccated F=Frozen L=Light Sensitive RT=Room Temperature]
Soluble: CHCl3, DMSO
Absorption: (in nm) 530
Emission: (in nm) 635
Extinction (103): 38
Molecular Formula: C20H18N2O2
CAS Number: [7385-67-3]
Alternative Name: Nile Blue A Oxazone
Unit Price (USD):
10 mg $58.28
Bulk Price/unit (when you buy 5 or more): $46.62
Description:
Lipophilic dye that stains intracellular lipid droplets to produce a bright red fluorescence in live cells.
Application:
Nile red is almost nonfluorescent in water but becomes intensely fluorescent, with a strong red emission when in a lipid-rich environment (EX552 nm/EM636 nm). But the fluorescence emission is environmentally sensitive. Intracellular fat vacuoles, filled with neutral lipids such as cholesterol, lipoproteins and triglycerides will fluoresce green (EX 485 nm/EM 525 nm) while polar lipids, such as phospholipids, will fluoresce red. The ratio of green:red fluorescence can be used to normalize cell size and dye uptake and to identify cells that have more "fat vacuoles". The probe can be added directly to the cell culture in order to label the fat vacuoles within adipocytes and these can then be analyzed by flow cytometry. Nile red has also been used to stain lipophilic proteins that have been separated by SDS-PAGE electrophoresis or to detect sphingolipids on thin-layer chromatograms.
For tissue staining, a stock solution of Nile red (500 ug/mL) in acetone is prepared and stored chilled and protected from light. A fresh staining solution of Nile red is made by the addition of 2-10 uL of the stock solution to 1 ml of 75% glycerol followed by brisk vortexing. The glycerol-dye solution is then briefly degassed by vacuum to remove bubbles. To stain frozen sections of tissue, a drop of the glycerol staining solution is added to each section and the preparation covered with a glass coverslip. After 5 min., the sections can be examined by fluorescence microscopy. Depending on the amount of tissue lipid present and the tissue thickness, the amount of Nile red required may need to be adjusted for optimum staining. Typically 1-5 ug dye/ml of 75% glycerol is required for maximum fluorescent staining.
References:
Fowler SD, Greenspan P (1985). "Application of Nile red, a fluorescent hydrophobic probe, for the detection of neutral lipid deposits in tissue sections: comparison with oil red O". Journal of Histochemistry and Cytochemistry 33(8): 833-836.
Makino A, Ishii K, Murate M, Hayakawa T, Suzuki Y, Suzuki M, Ito K, Fujisawa T, Matsuo H, Ishitsuka R, Kobayashi T (2006) "D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol alters cellular cholesterol homeostasis by modulating the endosome lipid domains." Biochemistry (45):14 4530-4541.
Werner T, Liebisch G, Grandl M, Schmitz G (2006) "Evaluation of a high-content screening fluorescence-based assay analyzing the pharmacological modulation of lipid homeostasis in human macrophages." Cytometry A (69):3 200-202.
Lamprecht A, Benoit JP (2003) "Simple liquid-chromatographic method for Nile Red quantification in cell culture in spite of photobleaching." J Chromatogr B Analyt Technol Biomed Life Sci. (787):2 415-419.
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