Robert,

What you are describing is typical of heat artifact.

I  bet the problem is the heat of the paraffin baths only because the problem is
also appearing in lymphoid tissue. 63 degrees is a little high for anything on
the tissue processor. Try using low melting point paraffin for infiltration on
the tissue processor.

I don't know if you have two processors but if you do, program one of them with
a shorter program for biopsies.  They do not require the same length of time in
wither the alcohols or paraffin baths as larger specimens do.  Tiny Biopsies
only need  20 minutes-1/2 hour in ea. paraffin bath.

I still bet the problem is the heat of the paraffin baths only because the
problem is also appearing in lymphoid tissue.

Also, check the heat the in oven you are using for drying the slides.  Try to
keep it a 59 degrees.

Hope these suggestions help.

Rande Kline, HT (ASCP)
Technical Services
EM Science/BDH




"SALA,ROBERT" <SALAR@siouxvalley.org> on 03/17/2000 11:34:27 AM

To:   'HistoNet' <histonet@pathology.swmed.edu>
cc:    (bcc: Rande Kline/EMI/Merck)
Subject:  Nuclear Staining



Please forgive me if I am going over old discussions.

We are having a problem with our nuclear staining.  It especially occurs on
tiny biopsies and on lymphoid tissue.  It is that hazy washed out nuclear
pattern.  We have tried about everything to correct the problem.  One our
paths is adamant that it is a heat artifact, I am leaning to incomplete
dehydration.  We have no heat above 63 C. in any of our procedure, we
dehydrate in ethanol and clear in a xylene substitute called Clear-Rite III.
This artifact in intermittent and does not affect all tissues on the run.
It appears to get worse towards the bottom of the slides.  VERY FRUSTRATING!

If anyone has any suggestions on how to correct this please let me know.
Thanks everyone.


Robert M. Sala M.T.(ASCP)
Histology Supervisor
Sioux Valley Hospital
1100 S. Euclid Ave.
Sioux Falls, SD  57117-5039