RE: [Histonet] Purple Haze.....

From:"Therersa Stegall"

Problem solved!  After considering amounts of Sta-on, gelatin, and using
a "naked" waterbath, all still produced the artifact.  It had to be the
hematoxylin, eh?  We used modified Harris, and considering the recipe
has acetic acid in it, thought that this was not the optimal for
decolorization in regressive staining.  Thought the haze could be an
indication of decaying hematoxylin; used HCl in our acid/water solution
this am, maybe it would help to capture the possibe alum hematein....
anyhow, it worked.  The slides are crisp and clear, free from artifact,
and now that the pathologists are happy, we are.......Thanks, Terre

>>> "Yaskovich, Ruth A (NIH/NIDCR)" 
10/08/03 01:48PM >>>
I used to get this problem once in awhile and haven't had it since
to charged slides. May-be in the amounts of gelatin? What kind of
hematoxylin are you using?
Ruth Yaskovich 

-----Original Message-----
From: Therersa Stegall [] 
Sent: Wednesday, October 08, 2003 1:50 PM
Subject: [Histonet] Purple Haze.....

   Thanks for naming the amorphous blight that has been plaguing us
This "Jimi Hendrix" artifact has been running 'round our brains for the
few weeks.  We have 1-started filtering the hematoxylin each morning,
2-alternated between Sta-on, gelatin, and naked waterbaths, 3-scrubbed
entire staining system, 4-decreased time in microwave, ovens, for
slides; and yet, we still find this on some biopsies.  Not always all
biopsies in a batch, this led us to ask GI about collection procedures,
possible air-drying, they insist it's not them.  We've checked pH on
accurately calibrated the bluing and acid/water washes (we stain
regressively), and just about run out of ideas.  I'm beginning to
the hematoxylin is leaving a precipitate, or maybe the microtomy is
inconsistent and ruining cell
integrity, or .... maybe it's terrorism!   Any ideas out there?

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