RE: [Histonet] Losing tissue on IHC slides

2013-07-29 Thread Sue Hunter
Good Morning
Another thing to try is different slides.  We have had issues with bad 
batches/lots of slides - even the Fisher Superfrost plus.  Especially if your 
purchasing department buys in quantity and then stores the slides in hot 
warehouses. This seems to  negate the charge on the slides - but in an 
inconsistent way.  You can have bad slides mixed with good ones in the same 
box.  Even if you don't want to change vendors, trying a different kind of 
charged slide would eliminate bad slides as part of the problem.  And just an 
FYI- we do our control slides the same way you do, (and use tap water) so I 
don't think that is your problem. We are using the "cheaper" Fisher Superfrost 
plus slides and they seem to work as well as the full priced ones.  We cut our 
immuno sections at 3um.
I know how frustrating this can be - I feel your pain!
Good luck!
Sue

Sue Hunter, Supervisor
Advanced Diagnostics
Beaumont Health System
Royal Oak MI
248-898-5146
shun...@beaumont.edu

 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Deloris Carter
Sent: Friday, July 26, 2013 5:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Losing tissue on IHC slides

Hi all,
I'm looking for some troubleshooting help.  In the last couple of months we've 
been having an increasing trend in losing tissue from our IHC slides.  We use a 
Ventana Benchmark XT.  PM is done on it quarterly.  The problem seems to be 
with the patient tissue, not the control tissue.  We use Fisherbrand Superfrost 
Plus slides, picking up the control by dipping the top end of the slide to 
avoid "double dipping".  We use tap water in the waterbaths which I know could 
cause an issue, but that's the way it's always been done here, so there's been 
no change to that part of the procedure. (We did try using DI water, but it 
didn't make any difference.
Tissue still fell off of slides.) We do use DI water to mix the bulk 
reagents.The control slides may sometimes have been cut for a longer than 
recommended time, but that part of the procedure hasn't changed either, same 
process as always.  If we run separate slides for control and patient tissue, 
we get much better results, but still have some tissue loss.  I can't say it's 
all of our tests, but it is a steadily worsening problem.
I'm going through tons of reagent, and I can't seem to nail down the problem.  
Ventana rep said to try DI water, don't double dip, try adhesive slides (which 
didn't really make any difference that we could tell).  I would appreciate any 
suggestions Deloris Carter Shawnee Mission Medical Center 
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RE: [Histonet] Losing tissue on IHC slides

2013-07-28 Thread Clare Thornton
I have a couple of things.  First off, are the sections coming off the slides 
entirely, or does it have a "chewed up" appearance?  If the sections are coming 
completely off, I would agree with the others that water is the issue.  Also, 
make sure you are cutting no thicker than 4 microns.  We had repeated a test 
twice due to sections falling off, and the tech who cut them has a tendency to 
cut on 5 microns.  I cut them the third time at 4 microns, and let the sections 
really flatten out on the water bath for a minute.  Then baked them well, and 
third time was the charm.  The sections remained on.  As far as the "double 
dipping" of precut control slides go, I spoke directly with a representative 
from Erie (the makers of all superfrost plus slides) and was told you could dip 
those slides a hundred times in water, it will not affect the charge on the 
slides.  We've never worried about double dipping and it has never caused us a 
problem.  If the sections have more of a chewed up appearance, then it's 
probably a combination of factors:  fatty tissue, not well fixed, strong 
antigen retrieval.  We've done something for years now that seems to help.  
After baking, we "post fix" some of our IHC slides in formalin for about 15 
minutes.  Rinse in tap water then put on stainer as usual.  This seems to help 
when certain antibodies like to chew up the tissue.  We don't do this will all 
antibodies, mostly breast markers and those with strong antigen retrieval.  I 
hope this helps!  Good luck.


Clare J. Thornton, HTL(ASCP)
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthorn...@dahlchase.com

From: histonet-boun...@lists.utsouthwestern.edu 
[histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Deloris Carter 
[dels...@gmail.com]
Sent: Friday, July 26, 2013 5:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Losing tissue on IHC slides

Hi all,
I'm looking for some troubleshooting help.  In the last couple of months
we've been having an increasing trend in losing tissue from our IHC
slides.  We use a Ventana Benchmark XT.  PM is done on it quarterly.  The
problem seems to be with the patient tissue, not the control tissue.  We
use Fisherbrand Superfrost Plus slides, picking up the control by dipping
the top end of the slide to avoid "double dipping".  We use tap water in
the waterbaths which I know could cause an issue, but that's the way it's
always been done here, so there's been no change to that part of the
procedure. (We did try using DI water, but it didn't make any difference.
Tissue still fell off of slides.) We do use DI water to mix the bulk
reagents.The control slides may sometimes have been cut for a longer than
recommended time, but that part of the procedure hasn't changed either,
same process as always.  If we run separate slides for control and patient
tissue, we get much better results, but still have some tissue loss.  I
can't say it's all of our tests, but it is a steadily worsening problem.
I'm going through tons of reagent, and I can't seem to nail down the
problem.  Ventana rep said to try DI water, don't double dip, try adhesive
slides (which didn't really make any difference that we could tell).  I
would appreciate any suggestions
Deloris Carter
Shawnee Mission Medical Center
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Re: [Histonet] Losing tissue on IHC slides

2013-07-27 Thread Benjamin
So the patient tissue and control are on the same slide, and the control is 
staying on but patient tissue is falling off? Maybe dont bake the control 
tissue just let them air dry, the humidity in the slide drying oven might be 
messing with the super frost plus slides. Its just a coating on the slide of 
some sort, i know after the first dip they dont work again like the first time, 
so just air dry the controls and bake after patient tissue is picked up.

Sent from my iPhone

On Jul 26, 2013, at 5:09 PM, Deloris Carter  wrote:

> Hi all,
> I'm looking for some troubleshooting help.  In the last couple of months
> we've been having an increasing trend in losing tissue from our IHC
> slides.  We use a Ventana Benchmark XT.  PM is done on it quarterly.  The
> problem seems to be with the patient tissue, not the control tissue.  We
> use Fisherbrand Superfrost Plus slides, picking up the control by dipping
> the top end of the slide to avoid "double dipping".  We use tap water in
> the waterbaths which I know could cause an issue, but that's the way it's
> always been done here, so there's been no change to that part of the
> procedure. (We did try using DI water, but it didn't make any difference.
> Tissue still fell off of slides.) We do use DI water to mix the bulk
> reagents.The control slides may sometimes have been cut for a longer than
> recommended time, but that part of the procedure hasn't changed either,
> same process as always.  If we run separate slides for control and patient
> tissue, we get much better results, but still have some tissue loss.  I
> can't say it's all of our tests, but it is a steadily worsening problem.
> I'm going through tons of reagent, and I can't seem to nail down the
> problem.  Ventana rep said to try DI water, don't double dip, try adhesive
> slides (which didn't really make any difference that we could tell).  I
> would appreciate any suggestions
> Deloris Carter
> Shawnee Mission Medical Center
> ___
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> Histonet@lists.utsouthwestern.edu
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Re: [Histonet] Losing tissue on IHC slides

2013-07-26 Thread Hans B Snyder
I agree water is always a potential issue.  Tissue can be finicky though.
 Have you tried poly-L-lysine coated slides?  They do help is some
occasions.  If you need a protocol, let me know, I can write one up, very
easy.

Hans B Snyder
Histologistics
60 Prescott Street
Worcester, MA 01605
508-308-7800
h...@histologistics.com 


On Fri, Jul 26, 2013 at 5:38 PM, Tony Auge  wrote:

> How are you drying your slides? If in an oven, what's the temp and
> duration? Try going longer and tap off excess water before drying.
>
>
>
> Tony Auge HTL (ASCP) QIHC
> Cell: (651) 373-4768
> Email: tony.a...@gmail.com
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Re: [Histonet] Losing tissue on IHC slides

2013-07-26 Thread Tony Auge
How are you drying your slides? If in an oven, what's the temp and
duration? Try going longer and tap off excess water before drying.



Tony Auge HTL (ASCP) QIHC
Cell: (651) 373-4768
Email: tony.a...@gmail.com
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