Hi all,
i am looking for a way to quantify the Alzarin red stain of my FFPE bone slices. My problem is that i want to compare the development of mineralization at specific time points and my original bones are all different in size. So in my opinin I have to account for the size differnece, but I have no idea how to analyze this. Furhtermore, I have a lot of slices to analyze and it would be great if there is maybe a program which runs semi-automatic (after I figured out which thresholds are best).
Thanks for your help!!
Vanessa
Hi Vanessa,
We are using image j software for quantifying alizarin red s staining on cell culture samples of differentiated cells. I think you can use same protocol as well. You can find protocols in researhgate. Also you can get quantitative results for alizarin red s by ELISA reading but its probably not suitable for tissue sections.
Hi Tayfun,
Thank you for your answer. Yes i already tried ImageJ and it works. The problem I have is that my bone sections differ in size (length and width of bone samples) and here I don't quite know how to account for that so it is comparable between different bones?
I used the technique on cell culture multidishes, so the seeded cell numbers and the surface area was all same which means there was no variable rather than calcium deposition sites. In your case sizes of the sections all differ, I assume. Maybe you should make a merged photo from whole section by photoshop first. Then calculate the surface area of each bone section. After that make the measurement with ImageJ and analyze the ratio of staining value with section size. If you make it for all your sections maybe you can get something solid to analyze and discuss. But as I told you, I have no experience on the issue, it's just a recommendation.
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