[Histonet] RE: Histonet Digest, Vol 102, Issue 24

[Joanne Clark]

We used to have this problem too, till we switched to Tru-Bond slides from Tru-Scientific.  Our contact is sara <@t> tru-scientific.com .  We also try to dry our slides for 1 hour in the 60 degree oven whenever possible and especially for really fatty tissues, like breast.  We haven't had any problems with antigen integrity.  I think greater than 60 could cook your tissue and affect things, but if you make sure it doesn't go any higher you should be OK.

Joanne Clark, HT
Histology Supervisor
Pathology Consultants of New Mexico

------------------------------

Message: 4
Date: Mon, 21 May 2012 09:25:21 -0500
From: "Brendal Finlay" <brendal.finlay <@t> medicalcenterclinic.com>
Subject: Re: [Histonet] Help
To: 'histonet <@t> lists.utsouthwestern.edu'
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <ea0c3755f29a151442a3fddc720e7d66 <@t> medicalcenterclinic.com>
Content-Type: text/plain; charset="utf-8"


Nancy,


We've had similar issues with fatty tissue falling off of the slides
while performing IHC.?? We use Superfrost + slides which we have found
to really hold the tissue well.?? Also, I have learned through reading
round on the??Histonet??that air drying doesn't completely remove the
water from the middle area of a tissue section.?? For this reason, we
no longer air dry at all unless it's a slide that was cut the day
before and just happened to be air dried.?? 


Our protocol changed to cutting the slides and draining them well,
then putting them in a 60 C oven for 15 minutes.????Then??the slides
are run??down to water on an automated stainer with another 15 minute
time in the oven on the stainer.?? 


A specific instance??when the tissue falls off,??was during antigen
retrieval in Trilogy in a pressure cooker.?? If the pressure was
manually released, this would cause the Trilogy to boil??and??it would
separate the tissue from the slide.?? Ourprotocol changed to 12
minutes in the pressure cooker with Trilogy, then around 8 minutes??to
wait for the pressure to release on it's own.?? We would then rinse
softly in distilled water to remove the??Trilogy.?? This also seemed
to help with the issue.

?? 

The combination of this??has worked??fairly well for us with some
exceptionally stubborn tissue still??attempting to fall off of the
slides.????I would love to??hear of other's experiences and??how they
resolved this.????


I do wonder about the length of time in your oven.?? I had spoken with
one of our Biocare reps about this when we encountered the problem and
he felt that longer than 30 minutes in the oven would damage the
specimen's IHC integrity.?? 


Brendal Finlay HT (ASCP)


----Original message-----
From: "Cloughley-Gray, Nancy" CloughleyN <@t> rvh.on.ca
Date: Fri, 18 May 2012 14:02:35 -0500
To:
"'histonet <@t> lists.utsouthwestern.edu'"histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Help

> I'm a Histotechnologist working in the Regional Hospital in Barrie,
ON Canada. We are using the Ventana Ultra for our Immunohistochemistry
(IHC). Since the end of February, we have been having issues with some
tissues lifting off our positive (marked with +) charged slides. It
seems to be mostly with the fatty and/or larger sections. We now dry
our slides for one hour at room temperature (R.T.) and an additional
hour at 60 degrees C. We cut our IHC sections at 4 um. Since we have
tried 2 different types of + slides and will be trying another type of
charged slide (from Newcomer this time) I was wondering if anyone has
any other suggestions?
> I also have another question regarding a QC (quality control) issue.
We use a multi-tissue control that is applied to the top of all our
test slides for IHC. One of our paths commented that there is some
positive staining in the smooth muscle nuclei of thenormal bowel when
we are testing for Progesterone (PR). We are using a Heat Induce
Epitope Retrieval (HEIR) of 36 minutes with CC1 (Ventana's proprietary
buffer @ pH of 8.0-8.5) and a primary antibody incubation time of 16
minutes with PR clone 1E2. (Ventana instrumentation provides
pre-diluted antibodies and the user adjusts the concentration of the
antibody by adjusting the time the primary antibody is incubated with
the tissue).
> I am concerned about the implications of this staining and I have
not been able to find a reference to this kind of unusual staining
pattern. The bowel tissue that we are using as QC is from a 62 year
old female patient. I was wondering if anyone has had any experience
with this kind of staining and /or any references that I could use.
> 
> Thanking you in advance,
> I look forward to your input,
> Nancy Cloughley-Gray MLT
>