Hi Mohammad,

Quantification of collagen can be done by hydroxyproline assay.

If you want to visualize the collagen, staining is an option. However, if the quantity of collagen is low, most of the antibodies may not give a strong enough signal. Since you are trying to look at collagen during degradation of the scaffold with time, this could be a problem. You could try SHG by two photon excitation if you have the scope for it.

Best,

Aja

Hi Mohammed,

There are lot of ways we can quantify collagen..

I would either adopt any colorimetry reactions or UV-Vis spectrophotometry.

If you have got UV-Vis spectrophotometer, then you can easily quantify  the collagen. Try to dissolve the pure collagen in the degradation solution, observe its UV-Vis absorption peak. Now make different concentrations of pure collagen in the degradation solution and plot a standard curve (conc Vs Abs).  Now after degradation study time points, you can collect the solution and record its UV-Vis absorption at particular wavelength and calculate the concentration.  Refer the below article for UV-VIs readings of collagen

http://www.sciencedirect.com/science/article/pii/S0147651309002826

If you wanna follow colorimetry reactions to quantify, you might look into this

https://www.researchgate.net/publication/233121162_Colorimetric_quantification_and_in_situ_detection_of_collagen

All the best..

Arun

Article Colorimetric quantification and in situ detection of collagen

Dear Aja and Arun

Thank you very much. I most read more about your suggestions :)