Abstract
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.
Highlights
Histology can be defined as the science that study the normal composition of tissues, while histological stains consider the key stone that used till to explain different component of tissues (1)
The original Masson's trichrome stain composed from four main stains, (i) Wiegert hematoxylin used to stain nucleus, (ii) Acid stain which target to stain muscles and red blood cells, (iii) Phosphatic acids that used to blech previous stains from collagen fibers and work as mordent to anionic dyes that stain collagen in step, and (iv) Collagen fiber stain that can be used to accomplish the goals of Masson's trichrome technique (3)
Routine and special stain The histopathological glass slides from previous steps were stain by routine Harris hematoxylin and alcoholic red eosin (3) in order to confirm classical lesions of chronic fasciolosis and fibrosis, the classical Masson's trichrome stain was used to stain slide which described by Layton and Bancroft (4) with Aniline Blue, slides with these stains considered as a control slides
Summary
Histology can be defined as the science that study the normal composition of tissues, while histological stains consider the key stone that used till to explain different component of tissues (1). Hematoxylin and Eosin stain (HE) and its modified protocols considered the main stain that applied routinely to visualize the components of tissues, in contrast, a special stain lead to expression a specific tissue component either in intracellular or extracellular location that could not clearly distinguished during HE routine staining (3). One of these components that need distinctive differentiation from surrounding cellular element is collagen fibers, these fibers are produced by mature fibrocytes and fibroblast in response to wide range of inflammatory cytokines produced by normal physiological and pathological process (4). Since this special stain contain many toxic even carcinogenic chemical materials in addition time consuming protocol, this study designed to eliminate these hazardous chemicals and decrease their concentrations, in addition, to improve the detection of collagen fibers using a new modified technique of Masson's trichrome stain
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