Trying to stain collagen by using just the aniline blue solution from a
trichrome method will not work. An acidified solution of aniline blue
will non-selectively stain all acidophilic tissues (red cells, muscle,
cytoplasm, collagen, etc.).
The reason that the trichrome methods distinguish collagen from other
tissue structures is that red cells and muscle have been pretreated with
phosphomolybdic acid prior to staining in aniline blue. The presence of
the aniline blue in those tissues inhibits (for a period of time) the
attachment of the blue dye. The principles of the trichrome techniques
depend on the molecular size of the dye molecules and the relative
porosity of the tissue proteins that are to be stained.
To selectively stain collagen without any other background staining, I
would suggest trying the following.
Bring the sections down to water as usual.
Treat the sections in 1% aqeous phosphomolybdic acid for 5 minutes
Drain off the acid. Do not wash as this will extract the PMA from the
Stain in acidified aniline blue (or any one of the blue or green stain
solutions used in a trichrome method) for 3 minutes.
Rinse very briefly in distilled water. Just a few seconds, no longer.
Dehydrate quickly through graded alcohols.
Clear in xylene and mount.
What I believe will happen is that the PMA will block entry of the dye
to all tissue structures except collagen, thus giving selective
coloration of the collagen fibres.
Jo-Ann Bader, Ms. wrote:
>Good Morning Everyone,
>I am looking for a stain for Collagen without a background stain. I have tried the Aniline blue from the Masson's Trichrome with unsatisfactory results.
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