[Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers

From:Guillermo Palao

Hello all:
  I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes.
  My protocol is as follows:
  1. Deparafinize and hydrate samples
  2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid
  3. Wash 2 min 0.01 N HCl
  4. Rinse in water
  5. Dehydrate and cover slides
  I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated.


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