Re: Returned mail: User unknown
here is a trichrome procedure i use for plastic sections, uses no phos.
acid
patsy
TRICHROME STAIN FOR GMA OR MMA EMBEDDED SAMPLES
Submitted by Patsy Ruegg, HT(ASCP)QIHC UCHSC Retired
Dry sections on 70dc hot plate for at least 20 minutes or 30 min
(62dc oven)
Mordant slides in saturated picric acid
Wash in running tap water 10
min
Rinse in deionized water
Stain in extra strength Weigert's - filtered 5 min
Rinse in running tap water 3-5
min
Stain in beibrich scarlet - acid fuchsin 30 min
Rinse quickly in deionized water
Stain in 2% lt. green in 2% acetic acid 10
min
Rinse in deionized water
3% acetic water for 4 to 5 min will enhance the blue
3% acetic in 95% alcohol, 2 changes, to remove stain from plastic media
Two changes of acetone
Clear in xylene and coverslip in permount
Results: Nuclei-black; mineralized bone-green; osteoid-red/orange;
collagen-green or blue (depending on counterstain); keratin, muscle, and
intracellular fibers - red
Biebrich scarlet-acid fuchsin
2% biebrich scarlet
1% acid fuchsin
Mix together:
45 ml bieb-scarlet (Can be reused but filter before each use)
5 ml acid fuchsin
Stir, then add 0.5 ml acetic acid
Extra Strength Weigert's
Sol A 10 gm Hematoxylin
100 ml 95% alcohol
Sol B 11.6 gm ferric chloride
980 ml deionized or distilled water
10 ml 25% HCL acid
Mix together and filter:
Sol A 5 ML
Sol B 25 ML
Abs. alcohol 20 ml
Aniline Blue
2.5 gm aniline blue
2 ml acetic acid
100 ml deionized or distilled water
2% Light Green
2 gm light green
98 ml distilled water
2 ml acetic acid
(Procedure orginally developed by Mabel Myli, Mayo Clinic many moons
ago)
Phi-Huynh Su wrote:
> Dear Histonet,
> 1. I have problems staining tendon with Masson Trichrome. It seems
> like parts of the fibers were dissolved by the method. I've stained
> same set of tendons with Alcian Blue, pH 2.5, and there was no obvious
> change. So I'd assume that acetic acid wasn't at fault. I'd think
> the phosphomolybdic acid was the culprit. I have repeated the
> procedure several times. When I stained femoral head sections with
> MT, it worked nicely. Please help. Is there any substitute for
> phosphomolybdic acid? Tissues were fixed in Bouin's solution. Since
> the tendons are quite thick, 5-8mm, they were fixed for 3 days. The
> excess picric acid was removed after rehydration of the tissues prior
> to hematoxylin step, not prior to processing.
>
> 2. What will happen if I forgot to shake the tissues for the first 12
> hours of fixation? (Bouin's fixative solution was used.)
>
> Thank you in advance
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