RE: [Histonet] aniline blue for trichrome

From:"Barry R Rittman"

Hi Liz
Technically methyl blue can be substituted for aniline blue WS. They are
both very acidic stains with sulfonate groups, similar absorption and
therefore similar color. The problem I had with methyl blue was that it
did not give as intense a color to connective tissue as the  aniline
blue WS.
I believe that the reason that methyl blue is substituted for aniline
blue WS is that it is easier to control the manufacturing process.
Aniline blue WS is a mixture of phenyl pararosanilines and I am told
that producing a consistent batch is difficult. I would recommend
looking at the original bottle that you had success with and not only
checking the CI but also the dye content and the batch number. If you
can get another bottle of the same then I would lay in a stock of this.
While suppliers don't like to do so they will often supply a dye of
specific batch number if you request this and if they have it in stock.
Wish I could suggest another solution.

-----Original Message-----
[] On Behalf Of
Elizabeth Chlipala
Sent: Wednesday, February 11, 2004 3:39 PM
To: 'HistoNet Server'
Subject: [Histonet] aniline blue for trichrome

Hello all
We are having a problem with our trichrome stain, we are not getting the
correct differentiation of muscle.  We normally don't have any problems
with this stain, since we always use fresh reagents.  We had an old
bottle of aniline blue from EMS that we have been using, I had ordered a
new bottle from Harleco they are both the same CI - 42755.  We started
having problems with the new aniline blue, but I'm not certain that this
is the reason why since what is not staining in the tissues is the
smooth muscle of vessels.  We have run this stain on rat and mouse
livers, both from carbon tetrachloride induced liver toxicity studies,
the morphological changes in this model include fibrosis, necrosis and
degenerating hepatocytes.  The rat livers look fine, except that the
smooth muscle of the vessels are not staining red.  The mouse livers
also have this problem, but in addition to that the degenerating
hepatocytes are staining slightly blue.  Also, when I checked into
ordering additional aniline blue from Sigma they use Methyl Blue, water
soluable - certified for use as Aniline Blue, but the CI is different
its 42780.  Any suggestions would be helpful.  
Thanks in advance
Elizabeth A. Chlipala, BS, HTL(ASCP)
Premier Histology Laboratory, LLC
P.O. Box 18592 
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
Ship to Address:
Premier Histology Laboratory
University of Colorado
MCBD, Room A3B40
Boulder, Colorado 80309
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