Hello everyone,

I am looking for a method to normalize alcian blue OD in micromasses, with the amount of cells. Normally cells are fixed, AB is extracted with GuHcl 8M, then OD is measured. At this point is there an easy way to evaluate cells number? I have tried SyBR green quantification, but together with GuHcl I have seen a 6 fold increase in the blank signal when I mix the two of them alone. Problem is that GuHcl is required. I have seen that neutralin red can be used on the same extracts (ref. http://ajpendo.physiology.org/content/299/2/E325.long#ref-29), but I could not find any specific protocol nor explanation. Also, I read that it's used with viable cells, but mine are fixed, so I was wondering whether someone could confirm or explain how it can be used as in the paper I linked.

I know that it's possible to extract GAGs in a cell Lysate, and from there quantify DNA as well, but it is more complex and longer as a protocol.

Thanks for any help!