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From:angelrj@email.uc.edu
To:histonet@Pathology.swmed.edu
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Date:Fri, 12 Mar 1999 14:05:40 -0500
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Thanks to all for the responses to my Masson's Trichrome question.  I
ran an experiment trying several different things.
1.  I used 2 different lots of Bouins.  I found no difference.
2.  I stained at different times in the Phosphomolybdic/Phosphotungstic
Acid.  At 5 min., 10 min., and 15 min.  I found no difference here
either.
3.  I rinsed 1 min. after the bouins instead of my usual 10 min.
4.  I took the actual temp. of the bouins solution, ensuring that it was
actually 56C.  Normally, I just put the container in the oven to preheat
for 1 1/2- 2 hours, just assuming it had reached the correct tempt.  (I
like to handle the bouins the least amount possible.)
5. I also used fresh bouins.  We normally reuse it about 5 times before
disposing of it.
6.  I read in our histology book that you should also rinse quickly when
dehydrating.  Alcohol also takes out some of the red stain.


The muscle on my slides turned out red with these test runs.  I'll have
to do more staining to check the consistency.  This stain was very dark
and bright.

My conclusion is:
1. I will only reuse the bouins one time, after filtering.  Then it will
be disposed of.
2. I will rinse only 1 minute after taking the slides out of the bouins.
3. I will leave the slides in the phosphotungstic/phosphomolybdic acid
10 min.
4. I will check the tempt. of the bouins to make sure it is the correct
tempt.
5. I will rinse quickly when dehydrating through the alcohols.

Thanks to everyone for their input.

Sincerely,

Rita Angel



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