Stain reagent kit BA4084A

for histopathology

Van Gieson stain is usually used for collagen fiber staining. Collagen fiber is distributed most widely and maximum among the three fibers. Principle: A double staining based on the different affinities of acid fuchsin and Picric acid towards collagen fiber and muscle fiber. Collagen fiber will bind acid fuchsin and become red to pink, while muscle fiber will bind Picric acid and appear yellow. Methods: 1. Fix tissues in 10% formaldehyde, hydrate in typical manner and embed. 2. Deparaffinize tissue in water. 3. Stain with Weigert iron hematoxylin for 5~10 minutes. Rinse in water. 4. Differentiate quickly in 1% hydrochloric acid ethanol. Rinse in running water for several minutes. 5. Stain in Van Gieson solution for 1~2 minutes. 6. Remove the stain and differentiate quickly in 95% ethanol for several seconds. 7. Dehydrate in absolute ethanol and clear in xylene. Mount with mounting media. Precaution: 1. Weigert’s Iron Hematoxylin solution is prepared by mixing equal parts of Solution A and Solution B prior to use. Do not mix beforehand or else will be oxidized and precipitated, resulting in the loss of staining capability. 2. Differentiate tissue quickly in 95% ethanol after VG Staining. 3. Because acid fuchsin is soluble in water, after staining in VG solution do not rinse in water. Instead, add 95% ethanol directly to differentiate and then dehydrate in absolute ethanol. Otherwise, the colors from the VG stain will weaken or discolor.