Greetings..We have finished reworking our 34BE12
keratin antibody from Dako.  All our biopsies are
fixed in 10% BNF and cut at 4 microns from a distilled
filled waterbath.  It is extremely important when
mounting sections on any kind of charged slides that
you slowly raise the slide out of the waterbath as you
let the water drain from under the tissue sections.
Trapped water will cause you nothing but heartache and
armache in its recutting.  Also, we have found
SurgiPath + slides and Shanndon + slides to be the
best so far.  We have had trouble with any slide made
by Erie which is sold may common vendors.  Ask who
their supplier is.  I have not tried Dakos slides yet
but have asked for a sample.  Next, dry slides until
tissues are white and place into the oven for 1 to 2
hours.  After deparaffinization, we steam sections in
Dako TRS for 20 minutes at temperatures of at least
95C (times should be adjusted for the temperature drop
when adding sections)  The total time at 95 C should
be at least 10 minutes (more is not good).  Then we
use PK at room temperature for 2 minutes.  Stain for
45 minutes (1-100) on the autostainer using Envision +
with 20 minutes in AEC.  The difference from just
steaming is amazing.  Let me know if you need any more
information.  Good luck.  pat

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