Carstairs Method for Fibrin and Platelets

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Platelet Pete
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Carstairs Method for Fibrin and Platelets

I was wondering if anyone has experience with the Carstairs differential stain. At the conclusion of staining, fibrin is supposed to stain bright red, platelets gray blue or navy, collagen bright blue and red cells clear yellow. I've tried to stain mouse small intestine, lung, liver, kidney, brain and spleen. I also tested on a clot that I made with mouse blood. There is one step in the protocol that reads "Differentiate with 1% phosphotungstic acid until the muscle is red and background is pale pink. That's a bit of a trick for me based on the above listed tissues and were fixed in formalin for >48 hours. Does anyone have any idea how long I should differentiate? In most cases, the red cells are yellow but are also occasionally purple to purplish blue. The collagen as described is bright blue, however, the platelets are not gray blue or navy, they are a medium blue and the fibrin is more of a fuchsia color than bright red. Any help would be appreciated. This is an awesome stain particularly for looking at platelets and fibrin in injury research.  Thanks, Platelet Pete