Oil Red

Oil-red-O is a lipid soluble lysochrome (fat stain) that has greater solubility in lipoid substances than in usual hydroalcoholic dye solvents. It is used to stain lipids, fatty acids, triglycerides, and cholesterol in frozen tissue samples, histological sections, and monolayer adipocyte cultures. It has very high depth of red color and yet leaves the cellular structures clear. In cell culture, Oil-red-O is most commonly used for visualization and quantification of adipogenic differentiation of mesenchymal stem cells in vitro. Oil droplets in mature adipocytes take up the dye and appear red in color

Specification

• Appearance: Red liquid
• Molecular weight: 408.49
• Molecular formula: C26H24N4O
• pH: pH 4.6 (yellow) to 6.0 (red)
• Cell Culture Tested
• Shelf life: 48 months
• Storage: 10°C-30°C
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How to Use

(1-Culture and treat cultured cells in tissue culture plate as needed (see other protocols

2Take the plate (35-mm) out of incubator and remove the medium

 3Add ~2 ml of PBS to wash the cells and remove PBS completely

4Add 2 ml of 10% formalin (room temperature) and incubate for 10 min at R

5Discard formalin and add 2 ml fresh formalin. Incubate for at least 1 hour, or longer (Cells can be kept in formalin for a couple of days before staining. Wrap with parafilm and cover with aluminum foil to prevent cells from drying)

6Remove formalin with a pipett

7Wash cells with 2 ml of ddH2O twic

8Wash cells with 2 ml of 60% isopropanol for 5 min at R

9Let the cells dry completely at RT. If possible, use a hairdryer to dr

10Add 1 ml of Oil Red O working solution and incubate at RT for 10 mi

11Remove Oil Red O solution and immediately add ddH2O. Wash the cells 4 times with ddH2

12Acquire images under the microscope for analysi

13Remove all the water and let dr

14Elute Oil Red O dye by adding 1 ml of 100% isopropanol and incubate for 10 min with gently shakin

15Pipet the isopropanol with Oil Red O up and down several times to ensure that all Oil Red O is in the solutio

16Transfer the solution to a 1.5-ml tub

17Measure OD at 500 nm using100% isopropanol as blan