[Histonet] Movats

[Betsy Molinari]

Hi Histonetters,
I have received several human vessels for paraffin processing and to stain the sections for H&E and Movats. The H&E were fine. The human sections turned brownish yellow with the Movats.The control which is canine small intestine was perfect.
The protocol is standard
Bouins 1hr in 58C waterbath
Rinse till yellow disappears
Rinse in DH2O
1% Alcian Blue -20 min
Rinse in running tap H2O -5min
Alkaline alcohol-1hr
Rinse 10 min tap H2O
Rinse in DH2O
Verhoff's Hematoxylin -15 min
3 changes DH2O
Differentiate in 2% FeCl
Rinse in DH2O
5% sodium Thiosulfate -1min
Rinse in running tap-10 min
Rinse in DH2O
Woodstain scarlet/acid fuchsin-1.5 min
Rinse in DH2O
Rinse in 0.5% acetic acid water
5% aqueous phosphotungstic acid -2 changes 5 min each
Rinse in 5% acetic acid water
Rinse in 3 changes absolute ETOH
6% alcoholic  Safran solution
Absolute alcohol-xylene-coverslip
The human slides were fine until the Safran step. When I removed them from the stain into the 100% they were a yellowish brown .Under the scope the colors were there, blue, red, yellow and black. But on the slide the tissue was that brownish yellow. The researcher does not like to strong yellow color. Since my control was fine I question if something was going on with their tissue. I do not know how the tissue was handled before it came into the lab. They were very calcified and were decaled for 1-3 days in Cal Rite. I do know they were not rinsed after decal and were put straight back into 10% NBF before I got them for processing.
Should I have used a human control instead of canine?  These were very large pieces that were crammed into the cassette.
Thanks for the help. Sorry for the long post.
Betsy Molinari HT(ASCP)
Texas Heart Institute
Cardiovascular Pathology
1101 Bates St.
Houston,TX
832-355=6524 (lab)
832-355-6812 (fax)



Betsy Molinari, HT (ASCP)
Sr. Histology Research Technician
CV Pathology Research

Texas Heart Institute
6770 Bertner Avenue, MC 1-283
Houston, TX 77030

Office: 832-355-6524 | Fax: 832-355-6812
Email: BMolinari at texasheart.org
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