Dear all,

This morning we performed an Alizarin Red Staining in human dermal fibroblasts. Although the staining itself did work (we saw red/orange colouring),  the colouring disappeared already after the first washing step.Thus,we were not able to quantify the amount of calcification.

I was wondering if anyone have had this problem as well, and know trics/solutions to overcome the problem of 'the dissapearing' colouring...

In addition, I used 4% formaldehyde (~10% formalin) to fixate the cells and the Alizarin Red had a pH of 4.2 (made less than a month ago).

Many thanks in regard,

Laura