Katrina's Collagen stain
(Modified Masson's Trichrome)

Application: For use as a stain in routine histology or a counterstain in IHC procedures using DAB. Works particularly well with beta catenin stained sections. Enabling the viewer to easily discriminate between epithelial and stromal components.  

Results:  Collagen - blue

Solutions List

5% Phosphotungtic Acid in DI water

Aniline Blue Solution
    2.5 g Aniline blue
    100 ml DI water
        2 ml Glacial Acetic Acid

1% Glacial Acetic Acid in DI water

Staining Procedure:

  1. Cut paraffin sections at 3m. and heat immobilze in the 80C oven for 30 min. 
  2. Deparaffinize and hydrate sections to DI water.
  3. Mordant in preheated Bouin's solution for 1 hr. at 56 C.
  4. Place sections in 5% Phosphotungtic Acid for 15 min. (discard sol'n)
  5. Place slides directly into Aniline Blue solution for 6 min. (save sol'n). 
  6. Rinse until clear in DI.
  7. Place in 1% Glacial Acetic acid for 4 min.
  8. Dehydrate quickly through 2 X 95% alcohol, 2 X of abs. alcohol, 6-8 dips each. Clear 3 X xylene and Mount.