staining

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From:Edna_J_Gonzalez/Powderject@powderject.com
To:vbaker60@yahoo.com, valleygal@aol.com, "HistoNet Server" <histonet@pathology.swmed.edu>
Reply-To:
Date:Tue, 28 Sep 1999 09:23:35 -0500
Content-Type:text/plain; charset=us-ascii



I was staining sucessfully my paraffin block with pig skin samples, the
color of the hamatoxylin and the eosin was perfect. I s\don't know what
happened but know is different. After staining in the hematoxylin, rinsing
in running tap water, I dip the slides two times in 1% HCl in 70% ethanol.
At this point the slides will turn light red or pink, but now they are not
turning to light red, they stay purple. I haven't change the procedure at
all. I just replaced the reagents with fresh ones. This hasn't happened
before to me. Can someone help me with this? Was is going on?

I was using the AFIP method:
2 changes in xylene for 2 minutes each
2 changes of absolute alcohol for 2 minutes each
2 changes of 95% ethanol for 2 minutes each
70% ethanol for 2 minutes
distilled water for 30 seconds
Harris' hematoxylin for 6 minutes
running tap  water for 2 minutes
2 dips in 1% HCl in 70% ethanol
running tap water for 1 minute
.3% ammonia water for 30 seconds
running tap water for 10 minutes
70% ethanol for 2 minutes
eosin for 1-2 minutes
2 changes of 95% ethanol for 2 minutes each
2 changes of absolute alcohol for 2 minutes each
2 changes in xylene for 2 minutes each.





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