Oil red O is a fat-soluble dye. Its solubility in lipids is greater than that in solvents. It is highly soluble in fats and can specifically stain neutral fats such as triglycerides in tissues. When tissue sections are placed in dye solution, the dye leaves the dye solution and dissolves in lipids (such as lipid droplets) inside the tissue, making the droplets appear orange. The oil red O dye produced by our company is used to prove the lipid content of tissue by the soluble property of dye.
Saturatued oil red O staining solution
Preparation of working solution:
For temporary use, the working solution should be prepared according to the ratio of saturated oil red O dye to distilled water = 6:4, thoroughly mixed and evenly, bathed in warm water at 60-70℃ for 30min, filtered after cooling, and prepared for use.
1．Take out frozen sections (the slides are generally stored at -20℃) and dry at room temperature.
2．Oil red staining: gently dip the slides into the oil red O working solution for 8-10min (cover with light protection).
3．Background differentiation: The slides were taken out, stayed for 3s and then immersed in two tanks of 60% isopropanol for differentiation, 3s and 5s respectively. The slides were immersed in 2 vats of pure water successively and washed for 10s each.
4．Hematoxylin staining: take out the section, stay for 3s, then immerse in haematoxylin and redye for 3-5min, soak in 3 vats of pure water, 5s, 10s, 30s, respectively.Hydrochloric acid alcohol differentiation, distilled water wash, blue liquid return blue, distilled water rinse.
5．Slides sealing: Glycerin gelatin adopted to seal slides.
6．Microscopic examination, image acquisition and analysis.
The lipid droplets are orange to red and the nuclei are blue.
1．If fresh tissue is frozen section, it needs to be fixed first and then stained; If the tissue is frozen section after fixation, the section can be dried and dyed directly.
2．The working solution must be ready for use. Heating process must be sealed, so as to avoid volatilization of isopropanol.
3．Pay attention to the action during the whole operation process, avoid fat loose or displacement.
4．The staining results can not be stored for a long time, and should be observed and photographed as soon as possible after sealing.
5．Glycerin gelatin is solidified at room temperature, and stored in an oven at 60℃ generally. If there are bubbles after sealing, the cover glass can not be pressed or forcibly pulled off the cover glass. The cover glass can be put into warm water, so that the cover glass can fall off by itself, and then resealing the cover glass to prevent fat displacement.
The liver tissue was stained with oil red O, the lipid was red, and the nucleus was blue.
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