Improving light microscopic detection of collagen by trichrome stain modification

In present study we aimed to introduce modifications in classical protocol applied to stain collagen fibers by Masson's trichrome stain, in order to decrease time and concentration of different steps and chemical in this traditional protocol. The experiment design contains a series of successive amendment steps based on previously successful steps, in which every step where be modified to get the best result, then the next step of staining protocol will be modified in the classical Masson's trichrome staining protocol in a samples of tissue from liver, heart and intestines of sheep diagnosed with chronic fasciolosis. The result showed that the new modified protocol gives the same staining efficiency to collagen fiber when we used Harris hematoxylin as a nuclear strainer or even excluded this stain from staining protocol. In conclusion this new modified staining protocol give a perfect staining reaction to collagen fibers in pathological samples which is similar to that obtained by Masson's trichrome stain, also this new modified procedure is less time consuming and less toxic effect on human and environment than other trichrome stains, and can be easily conducted either by the technician or the pathologists.