What is most effective way to stain, characterize, and analyze new collagen formation in remodeling tissue? I have seen trichrome and sirius red stains, curious if anyone has advice on which is best, or when to use one and not the other. Please share any other advice you may have on visualizing & characterizing newly created collagen using any techniques - histo, immunohistochemistry, PCR, etc. Thanks!
Safranine O/ fast green staining and Sirius red also can be used and you can quantitative estimation of collagen synthesis is also a method.
Masson's trichrome and Picrosirius (Sirius) red.
Evaluating picrosirius red should be by polarization (birefringence), because sirius red also binds to some components of basal membranes.
Just be aware of that.
Good luck
For staining, Masson's Trichrome. Go talk to the folks in Surgical Pathology and the Surg Path Histology lab at the med school. Also talk to some of the faculty in the Path department, there are experts in collagen, matrix and endothelial cells there.
Masson's Trichrome is the fastest and easiest approach. if you have a kit, you can do it by yourself. Sirius red is more difficult and you will need time to optimize it (depends from the tissue) but you would be able to quantify the birefringence signal, if you want. As bruce suggested talk to someone at Dept. of Pathology, they do Masson's trichrome on regular basis and probably can do it for you faster.
Masson's trichrome is very good. I did some quantification of collagen stained using Van Gieson's stain (which is like picro-sirius red) in mouse hearts. If you need good contrast, the Van Gieson is very quick because it's just one solution, and you will have red collagen fibres on a pale yellow background.
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