Re: [Histonet] IHC background

2022-09-29 Thread Rathborne, Toni via Histonet 

We have not encountered a covertile artifact with our Bond III, which we've had 
for over 10 years. It may be that the covertiles need to be replaced or cleaned 
more, but we have not experienced this. We also place a control tissue at the 
top of the slide, and often have large patient sections on the lower portion. 
Rene had some good suggestions. I would also call Leica to see if they can 
offer suggestions over the phone. Their technical support team has been very 
helpful over the years.
Best of luck.




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-Original Message-
From: Regan Fulton via Histonet 
Sent: Thursday, September 29, 2022 3:42 PM
To: RENE MORALES 
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC background


*** This is an External Email ***

Hi,

The artifact you are seeing is a well-known artifact of the Leica covertile
design.
It does limit the available footprint for placing samples.

Hope this helps.

Regan



Regan Fulton, M.D., Ph.D.
CEO and Co-Founder
Array Science, LLC
475 Gate 5 Road, #100
Sausalito, CA 94965
(415) 577-7360



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On Thu, Sep 29, 2022 at 12:37 PM RENE MORALES via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Paula,
> Regardless of the position of the tissue: control/ patient ‘s tissue, most
> likely is the  distribution of the reagents during staining, isn’t
> adequate. Syringe, staining  reagents expiration, calibration, etc.,   Have
> you contacted service from Leica? Hope the solution is found,
> Thanks,
>
>
> Sent from my iPhone
>
> > On Sep 29, 2022, at 12:00, Paula via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > Hello everyone,
> >
> >
> >
> > We have the Bond III and we ran an S100.  We placed 2 serial sections
> > vertically on the slide with the control tissue at the top for staining.
> The
> > patient's section at the top underneath the control tissue had background
> > staining, while the section on the bottom of that had no background
> > staining.  Can someone determine why this would happen?
> >
> >
> >
> > Thank you in advance.
> >
> > Paula
> >
> > Bio-Path Medical Group
> >
> > ___
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> > Histonet@lists.utsouthwestern.edu
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>
>
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Re: [Histonet] IHC background

2022-09-29 Thread Regan Fulton via Histonet 
Hi,

The artifact you are seeing is a well-known artifact of the Leica covertile
design.
It does limit the available footprint for placing samples.

Hope this helps.

Regan



Regan Fulton, M.D., Ph.D.
CEO and Co-Founder
Array Science, LLC
475 Gate 5 Road, #100
Sausalito, CA 94965
(415) 577-7360



www.arrayscience.com



On Thu, Sep 29, 2022 at 12:37 PM RENE MORALES via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Paula,
> Regardless of the position of the tissue: control/ patient ‘s tissue, most
> likely is the  distribution of the reagents during staining, isn’t
> adequate. Syringe, staining  reagents expiration, calibration, etc.,   Have
> you contacted service from Leica? Hope the solution is found,
> Thanks,
>
>
> Sent from my iPhone
>
> > On Sep 29, 2022, at 12:00, Paula via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > Hello everyone,
> >
> >
> >
> > We have the Bond III and we ran an S100.  We placed 2 serial sections
> > vertically on the slide with the control tissue at the top for staining.
> The
> > patient's section at the top underneath the control tissue had background
> > staining, while the section on the bottom of that had no background
> > staining.  Can someone determine why this would happen?
> >
> >
> >
> > Thank you in advance.
> >
> > Paula
> >
> > Bio-Path Medical Group
> >
> > ___
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ___
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>
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Re: [Histonet] IHC background

2022-09-29 Thread RENE MORALES via Histonet 
Paula,
Regardless of the position of the tissue: control/ patient ‘s tissue, most 
likely is the  distribution of the reagents during staining, isn’t adequate. 
Syringe, staining  reagents expiration, calibration, etc.,   Have you contacted 
service from Leica? Hope the solution is found,
Thanks,


Sent from my iPhone

> On Sep 29, 2022, at 12:00, Paula via Histonet 
>  wrote:
> 
> Hello everyone,
> 
> 
> 
> We have the Bond III and we ran an S100.  We placed 2 serial sections
> vertically on the slide with the control tissue at the top for staining. The
> patient's section at the top underneath the control tissue had background
> staining, while the section on the bottom of that had no background
> staining.  Can someone determine why this would happen?
> 
> 
> 
> Thank you in advance.
> 
> Paula
> 
> Bio-Path Medical Group
> 
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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[Histonet] IHC background

2022-09-29 Thread Paula via Histonet 
Hello everyone,

 

We have the Bond III and we ran an S100.  We placed 2 serial sections
vertically on the slide with the control tissue at the top for staining. The
patient's section at the top underneath the control tissue had background
staining, while the section on the bottom of that had no background
staining.  Can someone determine why this would happen?

 

Thank you in advance.

Paula

Bio-Path Medical Group

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Re: [Histonet] Epon Resin Embedding Question

2022-09-29 Thread Duraine, Lita R via Histonet 
Hi Brian,

I regularly use epon for TEM work.  Even though the instructions are good for 
some climates , epon is particular.
Here in Houston, I let my blocks cure for 5 to 7 days depending on how humid or 
rainy it is.  When I press a fingernail into the block , if it leaves a mark I 
know it needs a day or two more.  I generally cure at 62 degrees Celsius.

Dryer climates you can get away with shorter cure times.  Just do the 
fingernail test before starting to section.

If your blocks were in the oven for that many days already, then one of the 
components of the epon resin was no good or contaminated.  If that is so you 
may salvage your specimen by going backwards in a few steps in propylene oxide, 
or whatever other dehydration solvent you used before trying to cure in epon.  
After you get your sample loose then continue forward with new epon, ddsa, nma, 
and the harder dmp-30.

Regards,

Lita Duraine
Baylor College of Medicine
NRI TEM Core
1250 Moursund St.
Houston, Tx 77030




Sent from my Verizon, Samsung Galaxy smartphone


 Original message 
From: Paula Sicurello via Histonet 
Date: 9/28/22 10:29 PM (GMT-06:00)
To: "Cooper, Brian" , "Cooper, Brian via Histonet" 

Cc: "Navarro, Alexander" 
Subject: Re: [Histonet] Epon Resin Embedding Question

Hi Brian,Unfortunately, no.  The most you can do is to put it back in the oven 
and let it bake forva very long time.
You won't be able to section it.  It will gum up the knife edge.  If they are 
using a diamond knife they need to clean it right away.
To add insult to injury, the poorly polymerized blocks are a safety hazard.   
Epoxy resins are all toxic, carcinogenic, mutagenic, a reproductive hazard, or 
all of the above combined.  It’s only when the resin is fully polymerized that 
it becomes an inert plastic.
Perhaps someone else out in Histoland (Tim Morkin, are you out there?) has had 
luck dealing with blocks like those.
Sincerely,

Paula Sicurello

  On Wed, Sep 28, 2022 at 12:27 PM, Cooper, Brian via 
Histonet wrote:   Good afternoon Histonet,

Asking this question for a friend:  Is there a way to correct an improperly 
prepared Epon resin embedded block?  Following embedding, the block is soft and 
"tacky" to the touch.  This block is proving VERY difficult to section.  Any 
assistance from those of you doing EM in your labs would be greatly appreciated.

Thanks,

Brian D. Cooper, HT (ASCP)CMQIHCCM| Histology Supervisor
Department of Pathology and Laboratory Medicine
Children's Hospital Los Angeles
4650 Sunset Blvd MS#43- Los Angeles, CA 90027
Ph: 323.361.3357
bcoo...@chla.usc.edu

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