what is the measurement of Oil red O staining - (Apr/06/2010 )
I'm doing oil red O staining of macrophages,but what is the measurement of ORO staining?
The number of cells stained by ORO?
The fluorescense intensity of ORO?
Or comparing the red area in cells?
It all depends on what you are looking at... sections of tissue? plated cells?
Oil red O stains lipid droplets. What you measure, As Bob1 said, depends on tissues vs. cells. Assuming cells, you are probably best off taking a direct measure of ORO bound. To do this you need to make sure you wash all of the un-bound stain (background) from the cells. You can then elute the ORO in isoproanol, and quantify in a spectrophotometer; peak absorbance is 518nm, though we use a 492nm filter. For comparison between replicate experiments, you can formulate a standard curve of ORO diluted in Isopropyl alcohol.
Thank you, bob1 and Jah.
I am going to compare the degree of foam cell formation between 2 groups of mouse peritoneal macrophages.
As I know,there are 2 measurements for this:
1) ORO staining
2) Quantitation of intracellular contents of cholesterol
Are there any else ways?
For oil red O staining,is OD/mg protein suitable?
or fluorescence intensity?