I am currently examining osteogenic differentiation on titanium discs with surface coatings. In particular as the Ti discs are opaque and solid, we cannot use spectrophotometry to assess the cell samples without doing a cell scrape.
Many thanks for your help!
Himesh
Hi Himesh,
You can extract the Alizarin Red stain by using 10% 1-hexadecylpyridinium chloride and then quantify it by reading at 570nm. Use a small amount of the extraction solution to keep the color concentrated and dilute it down if it is too much. I would recommend doing replicates (minimum 3) if possible such that you can get an average/S.D.
There are some fluorescent dyes such as Calcein blue, xylene orange and Geimsa that will stain calcium deposition but they did not work very well for me compared to Alizarin Red staining/extraction. Alizarin Red actually fluoresces as well. I think that for these stains to work well, the calcium deposition needs to be concentrated in small areas and not diffuse.
Good Luck!
Tom
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