Hi, I am doing the experiments about the Osteogenic differentiation on hBMSC. I applied the Alazarin Red S staining for both control group and test group on 20th day. But the color of test group is light yellow. That should be the red-orange color on the Osteoblast. I could observe the morphological changes on cells. But I am really confused why the Osteoblast didn't show correct color by Alazarin Red S staining. I made Alazarin Red S solution several times and adjust the PH around 4.2 (4.1-4.3) within one month.

Anybody could help me to figure out what kind of issue caused light yellow COLOR for Osteoblast by Alazarin Red S staining?

Thanks very much

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