eosin fading

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
Date:Thu, 5 Aug 1999 10:29:35 -0600

One thing that is important is a good rinse after the bluing step, then going 
into 70% rinse,or the percentage of alcohol the eosin is dissolved in 
(sometimes not 70%) BEFORE going into the eosin y solution. Also make sure
the rinse after bluing solution is a minimum of 1 min, to wash away all
traces of the mild base used to "blue" the hematoxylin.  If one only does
a hand rinse instead of a running water rinse, you may want to consider
changing this water rinse EACH run, just to insure a correct rinse/pH 
adjustment, and prevent carryover/incorrect pH for the eosin..  

The rationale is to have the correct pH adjustment for the
eosin solution.  

Learned this from Jerry Fredenburgh in a staining workshop, and afterwards
noticed my eosin staining was more consistent, better. One tends to not
think about the importance of pH when doing rinse, blue, rinse then onto
the counterstaining portion of H&E staining.  Comes down to some chemical
rules! a forte of Jerry's.

Gayle Callis

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