Re: [Histonet] Ihc control tissues

[Colleen Forster via Histonet]

Charles,

I am in a research only lab as well. We create TMA's as control blocks
using the same idea that Val uses. Multiple tissues per block for all IHC
and also good for special stains. You can make nicely organized TMA's
manually very cost effectively as well. Feel free to reach out to me if
interested in more information.

Colleen Forster HT(ASCP)QIHC
cfors...@umn.edu

On Wed, May 8, 2024 at 9:56 AM Charles Riley via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Awesome thank you Val. I like the multi control option. Can save money in
> the end by using less reagents
>
> On Wed, May 8, 2024 at 10:52 AM Val T  wrote:
>
> > Hey Charles-
> >
> > Our research lab prepares a “multimouse” control. We embed 10 organs on
> > the same slide and it serves as a +/- control for just about every IHC
> > marker.
> > However, if you don’t want to do that- here are some tissues that would
> > make nice controls.
> >
> > > BCL-2- spleen, lymph nodes
> > > Caspase-3- spleen, lymph nodes
> > > CD117- skin, spleen, brain, lymph nodes
> > > CD62E (e-selectin), bone marrow, colon
> > > Collagen II, cartilage
> > > Desmin, heart, skeletal muscle
> > > GFAP, brain, spinal cord
> > > HMB45, skin
> > > Ki67, spleen, bowel
> > > p53, tumors, colon
> > > s100, colon, muscle
> >
> > Brain can serve as a negative control on just about all these. CD117 will
> > have some expression in the cerebellum, but will be negative in most
> other
> > areas.
> > Heart can be a negative control for GFAP.
> > Val
> >
> > > On May 8, 2024, at 6:59 AM, Charles Riley via Histonet <
> > histonet@lists.utsouthwestern.edu> wrote:
> > >
> > > Hello all,
> > >
> > > I am trying to put together a positive and negative tissue control list
> > for
> > > the following antibodies at my research core.  I came over from a
> > clinical
> > > pathology background and we used tonsils for a lot of these antibodies.
> > > Since rats and mice don't have tonsils I am looking for something they
> do
> > > have that can be used if possible to keep the controls as similar to
> the
> > > samples as possible.
> > >
> > > BCL-2
> > > Caspase-3
> > > CD117
> > > CD62E (e-selectin)
> > > Collagen II
> > > Desmin
> > > GFAP
> > > HMB45
> > > Ki67
> > > p53
> > > s100
> > > ___
> > > Histonet mailing list
> > > Histonet@lists.utsouthwestern.edu
> > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> ___
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>


-- 
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930
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Re: [Histonet] Ihc control tissues

[Charles Riley via Histonet]

Awesome thank you Val. I like the multi control option. Can save money in
the end by using less reagents

On Wed, May 8, 2024 at 10:52 AM Val T  wrote:

> Hey Charles-
>
> Our research lab prepares a “multimouse” control. We embed 10 organs on
> the same slide and it serves as a +/- control for just about every IHC
> marker.
> However, if you don’t want to do that- here are some tissues that would
> make nice controls.
>
> > BCL-2- spleen, lymph nodes
> > Caspase-3- spleen, lymph nodes
> > CD117- skin, spleen, brain, lymph nodes
> > CD62E (e-selectin), bone marrow, colon
> > Collagen II, cartilage
> > Desmin, heart, skeletal muscle
> > GFAP, brain, spinal cord
> > HMB45, skin
> > Ki67, spleen, bowel
> > p53, tumors, colon
> > s100, colon, muscle
>
> Brain can serve as a negative control on just about all these. CD117 will
> have some expression in the cerebellum, but will be negative in most other
> areas.
> Heart can be a negative control for GFAP.
> Val
>
> > On May 8, 2024, at 6:59 AM, Charles Riley via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > Hello all,
> >
> > I am trying to put together a positive and negative tissue control list
> for
> > the following antibodies at my research core.  I came over from a
> clinical
> > pathology background and we used tonsils for a lot of these antibodies.
> > Since rats and mice don't have tonsils I am looking for something they do
> > have that can be used if possible to keep the controls as similar to the
> > samples as possible.
> >
> > BCL-2
> > Caspase-3
> > CD117
> > CD62E (e-selectin)
> > Collagen II
> > Desmin
> > GFAP
> > HMB45
> > Ki67
> > p53
> > s100
> > ___
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Re: [Histonet] Ihc control tissues

[Val T via Histonet]

Hey Charles- 

Our research lab prepares a “multimouse” control. We embed 10 organs on the 
same slide and it serves as a +/- control for just about every IHC marker.
However, if you don’t want to do that- here are some tissues that would make 
nice controls.

> BCL-2- spleen, lymph nodes
> Caspase-3- spleen, lymph nodes
> CD117- skin, spleen, brain, lymph nodes
> CD62E (e-selectin), bone marrow, colon
> Collagen II, cartilage
> Desmin, heart, skeletal muscle
> GFAP, brain, spinal cord
> HMB45, skin
> Ki67, spleen, bowel
> p53, tumors, colon
> s100, colon, muscle

Brain can serve as a negative control on just about all these. CD117 will have 
some expression in the cerebellum, but will be negative in most other areas.
Heart can be a negative control for GFAP.
Val

> On May 8, 2024, at 6:59 AM, Charles Riley via Histonet 
>  wrote:
> 
> Hello all,
> 
> I am trying to put together a positive and negative tissue control list for
> the following antibodies at my research core.  I came over from a clinical
> pathology background and we used tonsils for a lot of these antibodies.
> Since rats and mice don't have tonsils I am looking for something they do
> have that can be used if possible to keep the controls as similar to the
> samples as possible.
> 
> BCL-2
> Caspase-3
> CD117
> CD62E (e-selectin)
> Collagen II
> Desmin
> GFAP
> HMB45
> Ki67
> p53
> s100
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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Re: [Histonet] IHC Validation Question

[Greg Dobbin via Histonet]

Hi Kara,
Use multi-tissue block controls with normal tissues that are treated
exactly the same (pre-analytically speaking) as your routine surgical
specimens. Consult NordiQC.org for recommended normal control tissue for
each IHC marker. The most used multi-tissue control block in my lab has
pieces of tonsil, pancreas, small bowel and liver.

*Greg Dobbin*
1205 Pleasant Grove Rd
Route 220
York,  PE  C0A 1P0
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Re: [Histonet] IHC gunk on slides

[Eddie Martin via Histonet]

Karen,

Regardless of the instrument you’re using, if related to a reagent, it may
be that your detection chromogen is breaking down and you need to report
it, or you have a material that is prepared onsite, and used onboard the
instrument, and unknowingly contaminated.

it may help if you can provide an image. It can be done with a mobile phone
up to the eyepiece if necessary.

Best wishes,
Eddie Martin
The National Institutes of Health
301-594-2054

On Thu, Mar 23, 2023 at 1:00 PM 
wrote:

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> Today's Topics:
>
>1. IHC gunk (Karen Heckford CA-San Francisco)
>2. Re: IHC gunk (Willis, Donna G)
>3. Re: IHC gunk (Thomas Podawiltz)
>4. Re: IHC gunk (Mac Donald, Jennifer)
>5. Re: Histonet Digest, Vol 232, Issue 7 (Eddie Martin)
>
>
>
> -- Forwarded message --
> From: Karen Heckford CA-San Francisco 
> To: Histonet 
> Cc:
> Bcc:
> Date: Wed, 22 Mar 2023 10:45:25 -0700
> Subject: [Histonet] IHC gunk
> Good Morning,
> I have recently developed a problem with contamination of some kind on my
> IHC slides.
> The contamination is black clumps and lays on top of the tissue.  I have
> been told it is bacteria but the Pathologist and I kind of doubt that.   I
> cannot get a good picture of it to show.  It may not be on all the slides
> on the same run or even the same antibody slide.
>
> I have decontaminated the whole system.  I have put fresh reagents on the
> instrument.   This stuff looks like it would wash off at the end of the run
> since it is sitting right on top of the tissue.  I do not see this stuff
> elsewhere on the slide, only the tissue.   Does not matter if the tissue
> was cut fresh or not.
>
> I have tried everything I can think of to get rid of it and still have this
> issue.  I use DiH20 from the hospital system.  Not sure if this may be the
> problem.  I can have Engineering test the DiH20.
>
> Any help would be greatly appreciated.
>
> Thanks,
>
> Karen Heckford HT ASCP CE
>
> Lead Histology Technician
>
> St. Mary's Medical Center
>
> 450 Stanyan St.
>
> San Francisco, Ca. 94117
>
> 415-750-5751
>
> karen.heckford@ commonspirit.org
>
>
>
>
> Caution:  This email message, including all content and attachments, is
> CONFIDENTIAL and may be of a nature that is LEGALLY PRIVILEGED.  The
> information contained in this email message is intended only for the use of
> the recipient(s) named above. If the reader of this message is not the
> intended recipient or an agent responsible for delivering it to the
> intended recipient, you have received this document in error.  Any further
> review, dissemination, distribution, or copying of this message is strictly
> prohibited.  If you have received this communication in error, please
> notify us  immediately by reply email.  Thank you
>
> Caution: This email is both proprietary and confidential, and not intended
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> you.
>
>
>
>
> -- Forwarded message --
> From: "Willis, Donna G" 
> To: Karen Heckford CA-San Francisco ,
> Histonet 
> Cc:
> Bcc:
> Date: Wed, 22 Mar 2023 17:57:09 +
> Subject: Re: [Histonet] IHC gunk
> What instrumentation are you using.  Have you talked to your vendor?
>
> Donna Willis
> Anatomic Pathology Manager
> Baylor Scott Health
> Baylor University Medical Center
> 3500 Gaston Ave|Dallas, Texas 75246
> 214-820-2465 office|214-725-6184 mobile
>
>
>
> -Original Message-
> From: Karen Heckford CA-San Francisco via Histonet <
> histonet@lists.utsouthwestern.edu>
> Sent: Wednesday, March 22, 2023 12:45 PM
> To: Histonet 
> Subject: {EXTERNAL} [Histonet] IHC gunk
>
>
> CAUTION:  This email originated outside of BSWH. The actual sender is  (
> histonet-boun...@lists.utsouthwestern.edu) which may be different from
> the display address in the From: field. Avoid action unless you know

Re: [Histonet] IHC gunk

[Mac Donald, Jennifer via Histonet]

Is it possible that it is hematoxylin precipitate?

-Original Message-
From: Karen Heckford CA-San Francisco via Histonet 
 
Sent: Wednesday, March 22, 2023 10:45 AM
To: Histonet 
Subject: [Histonet] IHC gunk

  EXTERNAL SENDER - Exercise caution with requests, links, and attachments.

Good Morning,
I have recently developed a problem with contamination of some kind on my IHC 
slides.
The contamination is black clumps and lays on top of the tissue.  I have
been told it is bacteria but the Pathologist and I kind of doubt that.   I
cannot get a good picture of it to show.  It may not be on all the slides on 
the same run or even the same antibody slide.

I have decontaminated the whole system.  I have put fresh reagents on the
instrument.   This stuff looks like it would wash off at the end of the run
since it is sitting right on top of the tissue.  I do not see this stuff
elsewhere on the slide, only the tissue.   Does not matter if the tissue
was cut fresh or not.

I have tried everything I can think of to get rid of it and still have this 
issue.  I use DiH20 from the hospital system.  Not sure if this may be the 
problem.  I can have Engineering test the DiH20.

Any help would be greatly appreciated.

Thanks,

Karen Heckford HT ASCP CE

Lead Histology Technician

St. Mary's Medical Center

450 Stanyan St.

San Francisco, Ca. 94117

415-750-5751

karen.heckford@ commonspirit.org




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Re: [Histonet] IHC gunk

[Thomas Podawiltz via Histonet]

Is it on all slides or just certain antibodies?


Sent from Yahoo Mail for iPhone


On Wednesday, March 22, 2023, 1:57 PM, Willis, Donna G via Histonet 
 wrote:

What instrumentation are you using.  Have you talked to your vendor?

Donna Willis
Anatomic Pathology Manager
Baylor Scott Health
Baylor University Medical Center
3500 Gaston Ave|Dallas, Texas 75246
214-820-2465 office|214-725-6184 mobile



-Original Message-
From: Karen Heckford CA-San Francisco via Histonet 
 
Sent: Wednesday, March 22, 2023 12:45 PM
To: Histonet 
Subject: {EXTERNAL} [Histonet] IHC gunk


CAUTION:  This email originated outside of BSWH. The actual sender is  
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Good Morning,
I have recently developed a problem with contamination of some kind on my IHC 
slides.
The contamination is black clumps and lays on top of the tissue.  I have
been told it is bacteria but the Pathologist and I kind of doubt that.  I
cannot get a good picture of it to show.  It may not be on all the slides on 
the same run or even the same antibody slide.

I have decontaminated the whole system.  I have put fresh reagents on the
instrument.  This stuff looks like it would wash off at the end of the run
since it is sitting right on top of the tissue.  I do not see this stuff
elsewhere on the slide, only the tissue.  Does not matter if the tissue
was cut fresh or not.

I have tried everything I can think of to get rid of it and still have this 
issue.  I use DiH20 from the hospital system.  Not sure if this may be the 
problem.  I can have Engineering test the DiH20.

Any help would be greatly appreciated.

Thanks,

Karen Heckford HT ASCP CE

Lead Histology Technician

St. Mary's Medical Center

450 Stanyan St.

San Francisco, Ca. 94117

415-750-5751

karen.heckford@ commonspirit.org




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Re: [Histonet] IHC gunk

[Willis, Donna G via Histonet]

What instrumentation are you using.  Have you talked to your vendor?

Donna Willis
Anatomic Pathology Manager
Baylor Scott Health
Baylor University Medical Center
3500 Gaston Ave|Dallas, Texas 75246
214-820-2465 office|214-725-6184 mobile



-Original Message-
From: Karen Heckford CA-San Francisco via Histonet 
 
Sent: Wednesday, March 22, 2023 12:45 PM
To: Histonet 
Subject: {EXTERNAL} [Histonet] IHC gunk


CAUTION:  This email originated outside of BSWH. The actual sender is  
(histonet-boun...@lists.utsouthwestern.edu) which may be different from the 
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safe. Report suspicious emails using the PhishAlarm button located in your 
Outlook ribbon.

Good Morning,
I have recently developed a problem with contamination of some kind on my IHC 
slides.
The contamination is black clumps and lays on top of the tissue.  I have
been told it is bacteria but the Pathologist and I kind of doubt that.   I
cannot get a good picture of it to show.  It may not be on all the slides on 
the same run or even the same antibody slide.

I have decontaminated the whole system.  I have put fresh reagents on the
instrument.   This stuff looks like it would wash off at the end of the run
since it is sitting right on top of the tissue.  I do not see this stuff
elsewhere on the slide, only the tissue.   Does not matter if the tissue
was cut fresh or not.

I have tried everything I can think of to get rid of it and still have this 
issue.  I use DiH20 from the hospital system.  Not sure if this may be the 
problem.  I can have Engineering test the DiH20.

Any help would be greatly appreciated.

Thanks,

Karen Heckford HT ASCP CE

Lead Histology Technician

St. Mary's Medical Center

450 Stanyan St.

San Francisco, Ca. 94117

415-750-5751

karen.heckford@ commonspirit.org




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Re: [Histonet] IHC staining of tendons and cartilage

[John Kiernan via Histonet]

You might like to look at this 1999 article from Microscopy Today, about 
keeping sections on slides.
https://publish.uwo.ca/~jkiernan/adhesivs.htm
John Kiernan
London, Canada.
= = =

From: Shirley A. Powell via Histonet 
Sent: March 21, 2023 2:56 PM
To: Charles Riley 
Cc: Histonet@lists.utsouthwestern.edu 
Subject: Re: [Histonet] IHC staining of tendons and cartilage

Hi Charles,

Shirley Powell here in humid Georgia.  I ran an IHC reference lab here for many 
years.  I had a problem with using charged slides for a lot of the tissues I 
processed.  I used manual and automation methods.  My tissues were washing off 
a lot.  I changed to an adhesive for the water bath called Sta-On and I think 
Surgipath was the company that made it.  Surgipath was bought out by Leica but 
they still sold it.  Sta-On was the best adhesive I had found and that worked 
for me for many years.  Whenever I do IHC that is what I use, especially bone, 
cartilage, bloody specimens, autopsy tissues, they stay on better.  Some other 
companies may be selling it now, like VWR/Avantor.  Just Google it.

Shirley

Shirley Powell, HTL(ASCP)
Technical Director Histology Curricular Support Laboratory
Pathology Department
Mercer University School of Medicine
powell...@mercer.edu
Phone: 478-301-2374
https://medicine.mercer.edu/

-Original Message-
From: Charles Riley via Histonet 
Sent: Tuesday, March 21, 2023 2:31 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC staining of tendons and cartilage

Hello all,

I am in a new position and it will potentially require doing a lot of IHC 
testing on cartilage and tendon samples. I have done some practice runs on my 
automated stainer and manually and am running into issues with the tissue 
sections falling off completely or folding over on itself during each process.

If anyone does staining like this routinely and has some pointers/tricks to try 
to get the samples to adhere to the slides better it would be greatly 
appreciated.

I have tried using charged slides from a variety of vendors and get similar 
results across the board.
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Re: [Histonet] IHC staining of tendons and cartilage

[Shirley A. Powell via Histonet]

Hi Charles, 

Shirley Powell here in humid Georgia.  I ran an IHC reference lab here for many 
years.  I had a problem with using charged slides for a lot of the tissues I 
processed.  I used manual and automation methods.  My tissues were washing off 
a lot.  I changed to an adhesive for the water bath called Sta-On and I think 
Surgipath was the company that made it.  Surgipath was bought out by Leica but 
they still sold it.  Sta-On was the best adhesive I had found and that worked 
for me for many years.  Whenever I do IHC that is what I use, especially bone, 
cartilage, bloody specimens, autopsy tissues, they stay on better.  Some other 
companies may be selling it now, like VWR/Avantor.  Just Google it. 
 
Shirley

Shirley Powell, HTL(ASCP)
Technical Director Histology Curricular Support Laboratory
Pathology Department
Mercer University School of Medicine
powell...@mercer.edu
Phone: 478-301-2374
https://medicine.mercer.edu/

-Original Message-
From: Charles Riley via Histonet  
Sent: Tuesday, March 21, 2023 2:31 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC staining of tendons and cartilage

Hello all,

I am in a new position and it will potentially require doing a lot of IHC 
testing on cartilage and tendon samples. I have done some practice runs on my 
automated stainer and manually and am running into issues with the tissue 
sections falling off completely or folding over on itself during each process.

If anyone does staining like this routinely and has some pointers/tricks to try 
to get the samples to adhere to the slides better it would be greatly 
appreciated.

I have tried using charged slides from a variety of vendors and get similar 
results across the board.
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Re: [Histonet] IHC background

[人生过客$ via Histonet]

你***


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Re: [Histonet] IHC background

[人生过客$ via Histonet]

？？？


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Re: [Histonet] IHC background

[人生过客$ via Histonet]

你理我不


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Re: [Histonet] IHC background

[Tony Henwood (SCHN) via Histonet]

Paula,

What does the artefact look like?

Another cause could be partial lifting of parts of the section, allowing 
reagents to become trapped under the section.
You commonly see it in thyroids where the colloid traps antibody and/or 
detection reagents under the colloid giving a particulate brown dust-like 
staining (sometimes looking like cocci).
You can also sometimes see it in fibrous tissue in sections.

Suggestions to solve this: check your sticky adhesive slides, or allow the 
slides to dry longer at room temperature before placing on the Bond (allows the 
adhesive time to work).

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children’s Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

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Re: [Histonet] IHC background

[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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>
>
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Re: [Histonet] IHC background

[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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Re: [Histonet] IHC background

[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
> 
> ___
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Re: [Histonet] IHC staining of cartilage

[Kurth, Virginia L via Histonet]

Make sure they sit at room temperature longer than normal and then dry them 
longer.  I found that cutting them thinner (3 microns) also helps. 



Ginny Kurth, HT, (ASCP)CM
UW Health Surgical Pathology
 Highland Ave
Madison, WI 






-Original Message-
From: Hobbs, Carl via Histonet  
Sent: Tuesday, March 01, 2022 1:30 PM
To: histonet 
Subject: Re: [Histonet] IHC staining of cartilage

WARNING: This email appears to have originated outside of the UW Health email 
system.
DO NOT CLICK on links or attachments unless you recognize the sender and know 
the content is safe.

Yes...a big problem for me, tooif you are using HIER??
Cartilage and bone are major tissue retention problems I cut Pwax sections of 
mouse long bones/knee joints for anti GFP IHC I still get a % fall off but,  
using manual HIER at 90C I manage to get enough sections to get good results 
However, I imagine that your autoIHC stainer prob uses 90C HIER?
I find Trajan 3 series slides more reliable than Superfrost Plus slides.
Sure, I need to dry my slides for 24 hrs in an operating fume hood ( we can't 
turn ours off/down) Then I place slides in a 60C oven for 2hrs.
Then dewax etc

Be interesting to read of other suggestions

Best wishes

Carl

Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge
Kings College London
London
SE1 1UL



020 7848 6810
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Re: [Histonet] IHC staining of cartilage

[Regan Fulton via Histonet]

Hi,

With regard to adhesion, we find TOMO slides are the best among many that
we have tried.

Interestingly, a creative histotech I once worked with used a diluted
Gorilla Glue smear on the slides just prior to picking up a cartilage
section.  It was tricky to get the section on while the glue was in a
workable condition, but the adhesion was phenomenally improved.  IHC worked
very well on these slides.

Best regards,

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 Tue, Mar 1, 2022 at 11:30 AM Hobbs, Carl via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Yes...a big problem for me, tooif you are using HIER??
> Cartilage and bone are major tissue retention problems
> I cut Pwax sections of mouse long bones/knee joints for anti GFP IHC
> I still get a % fall off but,  using manual HIER at 90C I manage to get
> enough sections to get good results
> However, I imagine that your autoIHC stainer prob uses 90C HIER?
> I find Trajan 3 series slides more reliable than Superfrost Plus slides.
> Sure, I need to dry my slides for 24 hrs in an operating fume hood ( we
> can't turn ours off/down)
> Then I place slides in a 60C oven for 2hrs.
> Then dewax etc
>
> Be interesting to read of other suggestions
>
> Best wishes
>
> Carl
>
> Carl Hobbs FIBMS
> Histology and Imaging Manager
> Wolfson CARD
> Guys Campus, London Bridge
> Kings College London
> London
> SE1 1UL
>
>
>
> 020 7848 6810
> ___
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Re: [Histonet] IHC staining of cartilage

[Hobbs, Carl via Histonet]

Yes...a big problem for me, tooif you are using HIER??
Cartilage and bone are major tissue retention problems 
I cut Pwax sections of mouse long bones/knee joints for anti GFP IHC
I still get a % fall off but,  using manual HIER at 90C I manage to get enough 
sections to get good results
However, I imagine that your autoIHC stainer prob uses 90C HIER?
I find Trajan 3 series slides more reliable than Superfrost Plus slides.
Sure, I need to dry my slides for 24 hrs in an operating fume hood ( we can't 
turn ours off/down)
Then I place slides in a 60C oven for 2hrs.
Then dewax etc

Be interesting to read of other suggestions

Best wishes

Carl

Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge 
Kings College London
London
SE1 1UL
 


020 7848 6810
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Re: [Histonet] IHC with zamboni's fixed tissues

[Tony Henwood (SCHN) via Histonet]

Hi Colleen,

I was involved in an Immunohistochemical study of the nerves in the human 
ureter that used Zamboni's fixative and cryostat sections. This study was 
before the invention of HIER.

If you are using Zamboni-fixed, paraffin embedded tissue then you will probably 
need to do HIER before many of the localisation antibody incubations since the 
fixative contains 2% formaldehyde. The same might be the case with some 
antibodies when applied to cryostat sections.

Edyvane, K. A., Trussell, D. C., Jonavicius, J., Henwood, A., & Marshall, V. R. 
(1992). Presence and regional variation in peptide-containing nerves in the 
human ureter. Journal of the autonomic nervous system, 39(2), 127-137.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal 
Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of 
Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead 
NSW 2145, AUSTRALIA 




-Original Message-
From: Colleen Forster via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Friday, 26 March 2021 9:42 AM
To: histonet-request 
Subject: [Histonet] IHC with zamboni's fixed tissues

Hello Histonet,

Can you give me feedback on trying to do IHC on samples fixed in Zamboni's 
fixative? I have an idea but need confirmation .

Thank you in advance

Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930
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Re: [Histonet] IHC validation after a new tissue processer is installed

[Greg Dobbin via Histonet]

Hi martha,
Prognostic markers must be re-validated (Eg.s Breast markers and CD117) as
you described.
Every Ab in your menu should be  tested (as you would for a new a new lot)
and do not forget to validate your H (with various tissue types) and SS
as well. For the H, if possible do side-by-side comparisons between old
and new processors (the downside of what is otherwise an exciting time!).
Greg

-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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Re: [Histonet] IHC validations

[Joe Myers via Histonet]

Dear Ms. Lucas:
Unfortunately, there is no specific CLIA standard that addresses the quantity 
of positive/negative specimens that should be tested during the procedure 
validation process.  As stated in the regulation excerpt shown below (section 
2), it is up to each laboratory to establish its own ‘performance 
specifications’ for procedures that ate developed ‘in-house’, as nearly all IHC 
procedures are.

42 CFR  493.1253 Standard: Establishment and verification of performance 
specifications.
(a) Applicability. Laboratories are not required to verify or establish 
performance specifications for any test system used by the laboratory before 
April 24, 2003.
(b)
(1) Verification of performance specifications. Each laboratory that introduces 
an unmodified, FDA-cleared or approved test system must do the following before 
reporting patient test results:
(i) Demonstrate that it can obtain performance specifications comparable to 
those established by the manufacturer for the following performance 
characteristics:
(A) Accuracy.
(B) Precision.
(C) Reportable range of test results for the test system.
(ii) Verify that the manufacturer's reference intervals (normal values) are 
appropriate for the laboratory's patient population.
(2) Establishment of performance specifications. Each laboratory that modifies 
an FDA-cleared or approved test system, or introduces a test system not subject 
to FDAclearance or approval (including methods developed in-house and 
standardized methods such as text book procedures), or uses a test system in 
which performance specifications are not provided by the manufacturer must, 
before reporting patienttest results, establish for each test system the 
performance specifications for the following performance characteristics, as 
applicable:
(i) Accuracy.
(ii) Precision.
(iii) Analytical sensitivity.
(iv) Analytical specificity to include interfering substances.
(v) Reportable range of test results for the test system.
(vi) Reference intervals (normal values).
(vii) Any other performance characteristic required for test performance.
(3) Determination of calibration and control procedures. The laboratory must 
determine the test system's calibration procedures and control procedures based 
upon the performance specifications verified or established under paragraph 
(b)(1) or (b)(2) of this section.

I hope you find this feedback useful.  I’ve prepared a document that outlines 
how a laboratory can comply with the standards and would be happy to share it 
with you; simply send me a message and I will respond accordingly.
Best Wishes,
Joe Myers, M.S., CT/QIHC(ASCP)

--

Message: 2
Date: Tue, 2 Jun 2020 09:25:03 -0700
From: "Paula" 
To: 
Subject: [Histonet] IHC validations

Hello,
I see 10 positive and 10 negative cases for CAP guidelines, but what about for 
CLIA?  What is their guideline to validate an IHC antibody?

Thanks in advance,
Paula

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Re: [Histonet] IHC validations

[Ingles Claire via Histonet]

Paula:
Usually CLIA follows CAP's lead or has less strict guidelines. Following the 
CAP guidelines should be good for both.
Claire

From: Paula via Histonet 
Sent: Tuesday, June 2, 2020 11:25 AM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] IHC validations

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Hello,

I see 10 positive and 10 negative cases for CAP guidelines, but what about
for CLIA?  What is their guideline to validate an IHC antibody?

Thanks in advance,

Paula

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Re: [Histonet] IHC validation

[Joe Myers via Histonet]

Kristy:
When validating an IHC procedure, regulatory guidelines like CLIA are not 
concerned so much with the types of specimens upon which the procedure will be 
applied as they are with how appropriately a given procedure detects different 
levels of protein expression, which, in turn, usually correlates to the  
‘degree of disease’.   In practical terms, this means that your lab’s efforts 
to validate should involve the staining of different tissues of the same type 
(i.e. skin), where the expression level ranges from ‘low‘ to ‘high’.  There are 
certainly a great deal more issues involved in procedure validation, but this 
is my attempt to answer your initial question.
Joe Myers, M.S., CT/QIHC(ASCP)

***

Message: 2
Date: Wed, 6 May 2020 05:43:22 -0700
From: Kristy Castillo 
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC validation

Would like to know when validating for IHC for a dermatology lab and just
for CLIA (no CAP), do you just need to show a shave, punch and excision 
lighting up?  Thanks!
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Re: [Histonet] IHC DAB

[Ana Maluenda via Histonet]

Hi Kristy,


As the others said, so many things that could be on the way...But making sure 
your system is based on a HRP enzyme is definitely a good start, as mentioned 
by Joe.


If you can share details, I might be able to help too.


Welcome to the IHC world!


Cheers,


Ana


Ana Maluenda
Research Assistant/Laboratory Manager
Atherothrombosis and Vascular Biology Laboratory



From: Joe Myers 
Sent: Saturday, 2 May 2020 4:49 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC DAB

Kristy:
I’m thinking along the same lines as Paula; is it possible that your detection 
reagent contains only Alkaline Phosphatase  (ALP) as there a reactive enzyme?  
If so, a peroxide/DAB solution simply won’t react with it.  Can’t wait to see 
your protocol, with detailed descriptions of the antigen, pretreatment reagent 
and ‘labeling’ enzyme.
Cheers,
Joe Myers, M.S.,CT/QIHC(ASCP)




From: Kristy Castillo via Histonet

 
Sent: Friday, May 1, 2020 11:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC DAB

Hi Histonetters,
We are starting our IHC (fun times), we are having trouble with the DAB 
lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our permanent 
red is working just fine.  Any ideas.
Thank you!
Kristy
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Re: [Histonet] IHC DAB

[Joe Myers via Histonet]

Kristy:
I’m thinking along the same lines as Paula; is it possible that your detection 
reagent contains only Alkaline Phosphatase  (ALP) as there a reactive enzyme?  
If so, a peroxide/DAB solution simply won’t react with it.  Can’t wait to see 
your protocol, with detailed descriptions of the antigen, pretreatment reagent 
and ‘labeling’ enzyme.
Cheers,
Joe Myers, M.S.,CT/QIHC(ASCP)




From: Kristy Castillo via Histonet

 
Sent: Friday, May 1, 2020 11:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC DAB

Hi Histonetters,
We are starting our IHC (fun times), we are having trouble with the DAB 
lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our permanent 
red is working just fine.  Any ideas.
Thank you!
Kristy
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Re: [Histonet] IHC DAB

[Paula Keene Pierce via Histonet]

Since you are newbies, I am just putting IHC 101 out there. 
Permanent red is for alkaline phosphatase detection and DAB is for HRP.
Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
Blue Lake DriveNorman, OK 73069PH 405-759-3953http://www.excaliburpathology.com

A sharp knife is nothing without a sharp eye. - Klingon Proverb 

On Friday, May 1, 2020, 10:29:00 AM CDT, Kristy Castillo via Histonet 
 wrote:  
 
 Hi Histonetters,

We are starting our IHC (fun times), we are having trouble with the DAB
lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our
permanent red is working just fine.  Any ideas.

Thank you!

Kristy
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Re: [Histonet] IHC DAB

[Kristy Castillo via Histonet]

Thanks for the tip!

On Fri, May 1, 2020 at 8:53 AM Val Tolley  wrote:

> It’s difficult to know without your full protocol, but you can easily test
> your DAB.
>
> Take a paper towel and add a drop of your secondary/HRP.  Add a drop of
> DAB to that.  If it doesn’t immediately turn brown, your hydrogen peroxide
> has likely degraded into water.  Make a fresh batch of DAB and try again.
>
>
>
> > On May 1, 2020, at 8:28 AM, Kristy Castillo via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > Hi Histonetters,
> >
> > We are starting our IHC (fun times), we are having trouble with the DAB
> > lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our
> > permanent red is working just fine.  Any ideas.
> >
> > Thank you!
> >
> > Kristy
> > ___
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Re: [Histonet] IHC DAB

[Val Tolley via Histonet]

It’s difficult to know without your full protocol, but you can easily test your 
DAB.

Take a paper towel and add a drop of your secondary/HRP.  Add a drop of DAB to 
that.  If it doesn’t immediately turn brown, your hydrogen peroxide has likely 
degraded into water.  Make a fresh batch of DAB and try again.



> On May 1, 2020, at 8:28 AM, Kristy Castillo via Histonet 
>  wrote:
> 
> Hi Histonetters,
> 
> We are starting our IHC (fun times), we are having trouble with the DAB
> lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our
> permanent red is working just fine.  Any ideas.
> 
> Thank you!
> 
> Kristy
> ___
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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Re: [Histonet] IHC DAB

[Porter, Amy via Histonet]

Hydrogen Peroxide gone bad / or not being added???


From: Kristy Castillo via Histonet 
Sent: Friday, May 1, 2020 11:00 AM
To: histonet@lists.utsouthwestern.edu 
Subject: [Histonet] IHC DAB

Hi Histonetters,

We are starting our IHC (fun times), we are having trouble with the DAB
lighting up.  Processed for 5 thru 10 minutes and still nothing.  Our
permanent red is working just fine.  Any ideas.

Thank you!

Kristy
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Re: [Histonet] IHC troubleshooting

[Greg Dobbin via Histonet]

Nancy,
You used the word “blot” in your reply to Charles, and while I can’t be
certain that you did not mean the same...I would like say in the interest
of clarity, that it is safer to carefully “wick” the water away from under
each section prior to baking.  Blotting (like we might do after a Gram
stain) would likely remove parts or all of the section.
All the best,
Greg
-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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Re: [Histonet] IHC troubleshooting

[Cartun, Richard via Histonet]

Are you using some form of antigen retrieval?  In addition to what has been 
discussed previously, antigen retrieval using heat (HIER) can cause tissue loss 
especially collagen.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 (Office)
(860) 545-2204 (Fax)
richard.car...@hhchealth.org

-Original Message-
From: Charles Riley via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, February 26, 2020 9:44 AM
To: Histo List
Subject: [Histonet] IHC troubleshooting

EXTERNAL email from Outside HHC! Do NOT open attachments or click links from 
unknown senders.

Our pathologists are complaining that chunks of the dermis are missing from IHC 
slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

--

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] IHC Troubleshooting

[Greg Dobbin via Histonet]

Hi Charles,
It adhesion problems can arise from several sources, some have already been
mentioned. Here are two that I have experienced:
1) Bad slides. Either a manufacturing defect such that the positive charge
is insufficient or the slides were somehow compromised after opening them
on the bench (perhaps exposed to moisture thus removing the charge).
Sometimes a new lot number be better but sometimes switching brands is the
quickest fix.
2) Ensuring all water is removed from under the section. I find that with
some charged slides (many in fact), water is trapped under the section and
does not drain out on standing and does not drain or evaporate when baking
at 60C. If the slide with trapped water is put on the instrument with
trapped water, the dewax step will lift much of the tissue.
Cheers,
-- 
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


*Everything in moderation...even moderation itself**!*
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Re: [Histonet] IHC troubleshooting

[Abdulbasit Andijany via Histonet]

In my experience;
If the the specimen is under-fixed or under-processed, the section will be 
sloughed off during IHC staining.

I find proper fixation time and drying the slides at room temperature (before 
baking them in oven) optimal.

Andy



Abdulbasit Andijany (Andy)
AMLS, BBMS, MSc, CT (ASCP)CM, HTL (ASCPI)
King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia


On 26 Feb 2020, at 18:35, John Garratt via Histonet 
mailto:histonet@lists.utsouthwestern.edu>> 
wrote:

Do you float and pick up your sections from a water bath of distilled water 
with no adhesive added? If not, doing that will help.

John

On Wed, Feb 26, 2020 at 6:41 AM, Charles Riley via Histonet 
mailto:histonet@lists.utsouthwestern.edu>> 
wrote:

Our pathologists are complaining that chunks of the dermis are missing from
IHC slides yet the entire section is present prior to staining.

Does anyone have any ideas what could cause the tissue to not adhere to the
slides throughout the staining process? We use the Leica Bond stainers.

--

Charles Riley BS HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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Re: [Histonet] IHC troubleshooting

[John Garratt via Histonet]

Do you float and pick up your sections from a water bath of distilled water 
with no adhesive added? If not, doing that will help.

John

On Wed, Feb 26, 2020 at 6:41 AM, Charles Riley via Histonet 
 wrote:

> Our pathologists are complaining that chunks of the dermis are missing from
> IHC slides yet the entire section is present prior to staining.
>
> Does anyone have any ideas what could cause the tissue to not adhere to the
> slides throughout the staining process? We use the Leica Bond stainers.
>
> --
>
> Charles Riley BS HT, HTL(ASCP)CM
>
> Histopathology Coordinator/ Mohs
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Re: [Histonet] IHC on Pig Tissues

[Liz Chlipala via Histonet]

Lori

You can't use a dual label detection system.  We have found most mouse polymers 
will cross react with porcine tissue.  We know that the dako one does.  You 
need to use the rabbit envision for rabbit monoclonal antibodies or with mouse 
monoclonals you need to use a rabbit anti-mouse and then a rabbit envision 
detection system.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

From: Lori Sorrells via Histonet 
Sent: Wednesday, January 29, 2020 1:35 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC on Pig Tissues

Good afternoon-
Has anyone run into problems when trying to perform IHC using rabbit mono or 
polyclonal antibodies on pig tissues? I typically do not have any issues with 
nonspecific staining or high background when using mouse monoclonal antibodies. 
I have used a variety of blockers including various sera in the diluents and 
blocking reagents. All samples are FFPE. I routinely use the DAKO EnVision+ 
Dual HRP system. Thanks in advance!

Lori Sorrells
Research Associate II
Revivicor, Inc.
1700 Kraft Drive, Suite 2400
Blacksburg, VA 24060

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Re: [Histonet] IHC mouse optomization

[Ana Maluenda via Histonet]

Hi Blanca,

It all depends on what you are trying to stain, how the tissue has been 
harvested/processed and what are your antibodies. If you are trying to use a 
primary mouse on mouse tissue, that is always tricky. You will need to probably 
explore your options with M.O.M kits. Otherwise, you may need to check your 
primary and secondaries. If you are using a secondary that is rat anti-mouse, 
you may also need to use an absorbed mouse secondary, to avoid too much 
non-specific staining.

Kind regards,

Ana

Ana Maluenda
Research Assistant/Laboratory Manager
Atherothrombosis and Vascular Biology Laboratory

Baker Heart and Diabetes Institute
75 Commercial Road, Melbourne VIC 3004
P (03) 8532 1359 E ana.malue...@baker.edu.au W www.baker.edu.au


-Original Message-
From: Margaryan, Naira 
Sent: Friday, 14 June 2019 3:27 AM
To: Blanca Lopez ; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC mouse optomization

Biocare Medical has pure ready to use secondary..

Naira

-Original Message-
From: Blanca Lopez 
Sent: Wednesday, June 12, 2019 10:57 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC mouse optomization

Morning,
I need a good advice in optimizing Mouse antibodies for research purposes. I 
use Dako/Agilent products. COX6B2 from Sigma is giving us hard times. Dako kit 
HRP has a mixture of mouse and rabbit so we think that might be the reason 
become negative.
Is there any tips for optimizing antibodies special for mouse tissue? Or if you 
can share your best procedures in IHC mouse stains. I can have a different ways 
and products to try. Any suggestion or opinions count. If you have any website 
that I can learn more about IHC for research mainly done in  Xenograft.
Thank you everybody:)

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu<mailto:blanca.lo...@utsouthwestern.edu>





UT Southwestern


Medical Center



The future of medicine, today.




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Re: [Histonet] IHC mouse optomization

[Margaryan, Naira via Histonet]

Biocare Medical has pure ready to use secondary.. 

Naira 

-Original Message-
From: Blanca Lopez  
Sent: Wednesday, June 12, 2019 10:57 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC mouse optomization

Morning,
I need a good advice in optimizing Mouse antibodies for research purposes. I 
use Dako/Agilent products. COX6B2 from Sigma is giving us hard times. Dako kit 
HRP has a mixture of mouse and rabbit so we think that might be the reason 
become negative.
Is there any tips for optimizing antibodies special for mouse tissue? Or if you 
can share your best procedures in IHC mouse stains. I can have a different ways 
and products to try. Any suggestion or opinions count. If you have any website 
that I can learn more about IHC for research mainly done in  Xenograft.
Thank you everybody:)

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu





UT Southwestern


Medical Center



The future of medicine, today.



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Re: [Histonet] IHC mouse optomization

[Colleen Forster via Histonet]

Biocare Medical Promark series are also great animal polymer based
detection that work very well.

https://biocare.net/products/detection/

Colleen Forster HT(ASCP)WQIUC
U of MN

On Wed, Jun 12, 2019 at 10:42 AM Paula Keene Pierce via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Vector Labs ImmPress kits rock for animal IHC.
> https://vectorlabs.com/browse/immpress-polymer-detection-kits-for-ihc
>
>
>
>
> Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830
> N Blue Lake DriveNorman, OK 73069PH 405-759-3953www.excaliburpathology.com
>
> On Wednesday, June 12, 2019, 10:13:48 AM CDT, Blanca Lopez via
> Histonet  wrote:
>
>  Morning,
> I need a good advice in optimizing Mouse antibodies for research purposes.
> I use Dako/Agilent products. COX6B2 from Sigma is giving us hard times.
> Dako kit HRP has a mixture of mouse and rabbit so we think that might be
> the reason become negative.
> Is there any tips for optimizing antibodies special for mouse tissue? Or
> if you can share your best procedures in IHC mouse stains. I can have a
> different ways and products to try. Any suggestion or opinions count. If
> you have any website that I can learn more about IHC for research mainly
> done in  Xenograft.
> Thank you everybody:)
>
> Blanca Lopez HT (ASCP)cm
> Senior Histotechnologist
> UT Southwestern Medical Center
> Harold C. Simmons Comprehensive Cancer Center
> UTSTR Biorepository Tissue Lab
> 6000 Harry Hines Blvd NB5.102
> Dallas, Texas 75390
> 214-648-7598
> blanca.lo...@utsouthwestern.edu
>
>
>
> 
>
> UT Southwestern
>
>
> Medical Center
>
>
>
> The future of medicine, today.
>
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-- 
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
B173 PWB  612-626-1930

*If submitting histology request please also forward to Lori Holm at
ho...@umn.edu *
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Re: [Histonet] IHC mouse optomization

[Paula Keene Pierce via Histonet]

Vector Labs ImmPress kits rock for animal IHC.
https://vectorlabs.com/browse/immpress-polymer-detection-kits-for-ihc




Paula Keene Pierce, BS, HTL(ASCP)HTPresidentExcalibur Pathology, Inc.5830 N 
Blue Lake DriveNorman, OK 73069PH 405-759-3953www.excaliburpathology.com 

On Wednesday, June 12, 2019, 10:13:48 AM CDT, Blanca Lopez via Histonet 
 wrote:  
 
 Morning,
I need a good advice in optimizing Mouse antibodies for research purposes. I 
use Dako/Agilent products. COX6B2 from Sigma is giving us hard times. Dako kit 
HRP has a mixture of mouse and rabbit so we think that might be the reason 
become negative.
Is there any tips for optimizing antibodies special for mouse tissue? Or if you 
can share your best procedures in IHC mouse stains. I can have a different ways 
and products to try. Any suggestion or opinions count. If you have any website 
that I can learn more about IHC for research mainly done in  Xenograft.
Thank you everybody:)

Blanca Lopez HT (ASCP)cm
Senior Histotechnologist
UT Southwestern Medical Center
Harold C. Simmons Comprehensive Cancer Center
UTSTR Biorepository Tissue Lab
6000 Harry Hines Blvd NB5.102
Dallas, Texas 75390
214-648-7598
blanca.lo...@utsouthwestern.edu





UT Southwestern


Medical Center



The future of medicine, today.

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Re: [Histonet] IHC Control Slide Longevity

[Jan Shivers via Histonet]

Hi Sandy,

We cut CD31, CD45, CD45RA, CD117, CD204, and MUM1 controls fresh (same
morning as staining; case material as well).

We've never had any problems with Synaptophysin; it's always worked very
well on unstained controls that are up to a year old (stored in the fridge).

Best regards,
Jan

Jan Shivers
Senior Scientist
IHC/Histology Section Manager
Pathology Teaching Program
Veterinary Diagnostic Laboratory
University of Minnesota
1333 Gortner Ave.
St. Paul, MN  55108
612-624-7297
shive...@umn.edu

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On Thu, Feb 28, 2019 at 12:57 PM Sandra Cheasty via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hello all,
> Do any of you have to cut any of you antibody controls
> "fresh"? We currently cut the MUM-1 fresh, and the Synaptophysin. (We
> seldom do a Syanpt., and have found the fresh cut patient tissue has been
> positive while the not-so-fresh positive control slide is negative.
> Any comments are - as always, are appreciated.
> Cheers,
> Sandy
>
> Sandra J. Cheasty, HT (ASCP)
> Histology & Necropsy Supervisor
> UW-Madison, School of Veterinary Medicine
>
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Re: [Histonet] IHC Billing - 88342 vs. 88341

[Perl , Alison via Histonet]

Morning everyone
I got a couple of inquiries about my LIS. We use IntelliPath from NetSoft; 
we're a physician-owned lab with separate clinical and pathology labs. They 
were able to customize logic for us to make sure IHC and SS get billed 
correctly without manual intervention 99% of the time. We're working on an 
interface, but it currently just exports to an Excel file that we send to 
Billing. 

Hope that helps! If you have other questions just email me

Alison Perl, HTL(ASCP)CM
Anatomic Pathology Manager 
CareMount Medical
(914) 302-8424
ap...@cmmedical.com


-Original Message-
From: Cartun, Richard via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Monday, January 21, 2019 3:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL] [Histonet] IHC Billing - 88342 vs. 88341

For those of you doing clinical IHC testing, have you been able to automate the 
billing of IHC (88342 vs. 88341) or do you still manual bill?  Our IT staff 
wants to build-in a "88342" and a ""88341" for every antibody that we run and 
then have the pathologist select the appropriate one for billing.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 (Office)
(860) 545-2204 (Fax)
richard.car...@hhchealth.org


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Re: [Histonet] IHC Billing - 88342 vs. 88341

[Perl , Alison via Histonet]

Hi Dr Cartun
That was our worst-case scenario, but our LIS was able to build logic so that 
it will convert 88342 to 88341 after the 1st, for each specimen (rather than 
case). It's been working well for the past 6 months or so...

Alison Perl, HTL(ASCP)CM
Anatomic Pathology Manager 
CareMount Medical
(914) 302-8424
ap...@cmmedical.com


-Original Message-
From: Cartun, Richard via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Monday, January 21, 2019 3:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: [EXTERNAL] [Histonet] IHC Billing - 88342 vs. 88341

For those of you doing clinical IHC testing, have you been able to automate the 
billing of IHC (88342 vs. 88341) or do you still manual bill?  Our IT staff 
wants to build-in a "88342" and a ""88341" for every antibody that we run and 
then have the pathologist select the appropriate one for billing.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology/Morphologic 
Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 (Office)
(860) 545-2204 (Fax)
richard.car...@hhchealth.org


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Re: [Histonet] IHC-H HELP...

[Greg Dobbin via Histonet]

Hi Curt,
You can de-stain the hematoxylin counter stain and then do H and all
should be fine. Subsequent IHC is possible but obviously you would need a
different color chromogen to differentiate the new stain from the previous
one.

There may be a problem  getting the section stay on through a second IHC
procedure but maybe not. I would stain a control section for the desired Ab
using the retrieval method that was used initially and see how the control
looks. If the pathologist thinks the that this control is adequate for
interpretation, then restain the slides with the desired Ab and no
additional retrieval.

Cheers,
Greg

-- 
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1205 Pleasant Grove Rd
RR#2 York,
PE  C0A 1P0


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Re: [Histonet] IHC-H HELP...

[Tony Henwood (SCHN) via Histonet]

Yep,

Definitely an issue.

You can easily stain the IPX slides with H, though the discernibility of the 
cells and tissue structure with the H will depend on the degree of DAB 
product laid down (eg I would expect it to be difficult with a Vimentin IPX 
compared to a CD15).

(Grosset, A. A., Loayza-Vega, K., Adam-Granger, É., Birlea, M., Gilks, B., 
Nguyen, B., ... & Trudel, D. (2017). Hematoxylin and Eosin Counterstaining 
Protocol for Immunohistochemistry Interpretation and Diagnosis. Applied 
immunohistochemistry & molecular morphology: AIMM.)

As for doing another IPX on the existing IPX stained section (with DAB as the 
chromogen), you will have to use a different label (eg alkaline phosphatase). 
The result will depend on the cell compartment the two antigens exist. If the 
DAB is nuclear, then a cytoplasm or cytoplasmic membrane localisation with the 
Alk Phosphatase will work. If both antigens are cytoplasmic, then you will not 
see co-localisation in the same cell since the DAB will prevent any antibody 
binding in the same compartment.

Assuming the above is good, then since the antigen retrieval tends to reverse 
over time, I would include a short retrieval before the second antibody.

Now after all that, I hope the section stays on the slide!


Regards 

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-Original Message-
From: Curt via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, 4 January 2019 4:05 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC-H HELP...

Need help before I turn a mistake into an irreparable mistake...

We have some unstained slides that were supposed to get stained with H but my 
guy stained them with IHC. It's complicated, we received slides and a block, 
the block was for IHC, the unstained slides were for H, he inverted the 
process) The point is, now the unstained slides are stained with IHC... I know 
we cannot destain the IHC but we can simply run and H over them... the real 
question I have is subsequent to the H this pathologist generally likes to 
see the H then order IHC on them based on what he sees (we only have these 
few unstained slides, don't have blocks to recut)...

So the question is... if we've already run IHC, then followed that with and 
H, can we return to run IHC on the slides again? would you want to skip any 
pre-treatment, antigen retrieval

I don't see this working too well myself, if they're already stained with DAB, 
that would be present on the second stain...

Thoughts?

Thanks for your help.

Curt

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Re: [Histonet] IHC Negative reagent controls

[John Garratt via Histonet]

The article referenced below may be of interest.

Standardization of Negative Controls in Diagnostic Immunohistochemistry: 
Recommendations From the International Ad Hoc Expert Panel
https://www.researchgate.net/publication/261516067_Standardization_of_Negative_Controls_in_Diagnostic_Immunohistochemistry_Recommendations_From_the_International_Ad_Hoc_Expert_Panel


John

www.ciqc.ca

‐‐‐ Original Message ‐‐‐
On Wednesday, January 2, 2019 6:48 PM, Cayman Fleck via Histonet 
 wrote:

> A question that came up regarding negative reagent controls for 
> IHC...currently using Ventana i-View. Our regular negative control goes 
> through the standard antigen retrieval steps, like 99% of our antibodies. 
> However there are a small number of antibodies that require enzyme as well 
> (Protease 1). I've seen a number of suggestions regarding this for the 
> negative reagent control...some say use an additional negative control 
> protocol that includes the protease, some say to use a single negative 
> control protocol and just include the harshest cell conditioning that any of 
> your protocols use (so basically use the cell conditioning + protease 
> negative control for all antibodies)...i-View is not polymer-based so we need 
> to continue using negative controls. Any thoughts or advice?
>
> Frank
>
> Sent from Outlookhttp://aka.ms/weboutlook
>
> Histonet mailing list
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Re: [Histonet] IHC Negative reagent controls

[Tony Henwood (SCHN) via Histonet]

Hi Cayman,

Unfortunately, applying HIER to a negative control for an antibody that 
requires enzyme retrieval (or no retrieval at all) is not appropriate.
The pre-treatment processes are different and could unmask different epitopes.
If you are using a negative control then the whole procedure needs to be same 
with the exception or replacing the localisation antibody with an Isotypic 
antibody solution. (Isotype controls are primary antibodies that lack 
specificity to the target, but match the class and type of the primary antibody 
used in the application.)

For example, applying citrate or EDTA HIER to sections prior to using the CD99 
antibody (clone 12E7) can reveal perinuclear (golgi-like) staining of some 
tumours (eg some colonic carcinomas) but this is not seen if enzyme retrieval 
is used.

Hope this is useful

Regards 

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-Original Message-
From: Cayman Fleck via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, 3 January 2019 1:48 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Negative reagent controls

A question that came up regarding negative reagent controls for IHC...currently 
using Ventana i-View.  Our regular negative control goes through the standard 
antigen retrieval steps, like 99% of our antibodies.  However there are a small 
number of antibodies that require enzyme as well (Protease 1).  I've seen a 
number of suggestions regarding this for the negative reagent control...some 
say use an additional negative control protocol that includes the protease, 
some say to use a single negative control protocol and just include the 
harshest cell conditioning that any of your protocols use (so basically use the 
cell conditioning + protease negative control for all antibodies)...i-View is 
not polymer-based so we need to continue using negative controls.  Any thoughts 
or advice?

Frank

Sent from Outlook 
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Re: [Histonet] IHC Stainer

[Colleen Forster via Histonet]

I am with Jennifer on the IntelliPath.I have just gotten 2 of them in
my lab.

The new instrument was demoed at NSH last year. It is an amazing
machine,I wish I could have had that one VEry limited numbers will be
avaiable in November with the main stream sales beginning in January.

Colleen Forster HT(ASCP)QIHC
U of MN


On Thu, Aug 30, 2018 at 12:58 PM, Jennifer Kempf via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Biocare Medical Intellipath...its completely open and is a continuous load
> machine, meaning you can start a run on one rack while still loading
> programs on the other racks if you choose.  Biocare is coming out with a
> new machine in January (I think), that is supposed to do heat retrieval
> online as well, instead of in the pressure cooker, but I haven't seen it in
> action yet.
>
> -Original Message-
> From: Sandra Cheasty [mailto:sandra.chea...@wisc.edu]
> Sent: Thursday, August 30, 2018 9:40 AM
> To: Histonet (histonet@lists.utsouthwestern.edu)  utsouthwestern.edu>
> Subject: [Histonet] IHC Stainer
>
> Hello all,
> We are in the market for a new IHC stainer. (Our Lab
> Vision 720 is no longer reliable.) We average about 20 slides a day, but
> sometimes get research projects that are 50+ slides.  Not interested in a
> closed system; it is for a veterinary histology lab, and we need the
> flexibility to tweak antibodies for dogs, cats, horses, and zoo animals.
> I'd really appreciate feedback from users who are in a similar lab setting.
>
> Cheers!
> Sandy
>
> Sandra J. Cheasty, HT (ASCP)
> Histology & Necropsy Supervisor
> UW-Madison, School of Veterinary Medicine
>
>
>
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>



-- 
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
B173 PWB  612-626-1930

*If submitting histology request please also forward to Lori Holm at
ho...@umn.edu *
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Re: [Histonet] IHC Stainer

[Jennifer Kempf via Histonet]

Biocare Medical Intellipath...its completely open and is a continuous load 
machine, meaning you can start a run on one rack while still loading programs 
on the other racks if you choose.  Biocare is coming out with a new machine in 
January (I think), that is supposed to do heat retrieval online as well, 
instead of in the pressure cooker, but I haven't seen it in action yet. 

-Original Message-
From: Sandra Cheasty [mailto:sandra.chea...@wisc.edu] 
Sent: Thursday, August 30, 2018 9:40 AM
To: Histonet (histonet@lists.utsouthwestern.edu) 

Subject: [Histonet] IHC Stainer

Hello all,
We are in the market for a new IHC stainer. (Our Lab Vision 720 
is no longer reliable.) We average about 20 slides a day, but sometimes get 
research projects that are 50+ slides.  Not interested in a closed system; it 
is for a veterinary histology lab, and we need the flexibility to tweak 
antibodies for dogs, cats, horses, and zoo animals. I'd really appreciate 
feedback from users who are in a similar lab setting.

Cheers!
Sandy

Sandra J. Cheasty, HT (ASCP)
Histology & Necropsy Supervisor
UW-Madison, School of Veterinary Medicine



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Re: [Histonet] IHC Stainer

[Rene J Buesa via Histonet]

Get a DAKO IHC stainer (even if it is "refurbished")René 

On Thursday, August 30, 2018 9:56 AM, Sandra Cheasty via Histonet 
 wrote:
 

 Hello all,
                We are in the market for a new IHC stainer. (Our Lab Vision 720 
is no longer reliable.) We average about 20 slides a day, but sometimes get 
research projects that are 50+ slides.  Not interested in a closed system; it 
is for a veterinary histology lab, and we need the flexibility to tweak 
antibodies for dogs, cats, horses, and zoo animals. I'd really appreciate 
feedback from users who are in a similar lab setting.

Cheers!
Sandy

Sandra J. Cheasty, HT (ASCP)
Histology & Necropsy Supervisor
UW-Madison, School of Veterinary Medicine

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Re: [Histonet] IHC System from StatLab

[Jeanine Ronkowski via Histonet]

All:For those of you who saw my posting last week and we’re wondering about the 
outcome, I’m sorry to say that I’ve received very little useful feedback.  Two 
individuals from StatLab sent me separate, cryptic messages indicating that 
they would be be happy to either meet with me or provide a demo of this unnamed 
instrument, but neither have provided me with a brochure or any other 
documentation, as I requested in my response.
I’ve never done this before, so I would like to ask this group – Is this 
behavior normal?  I’m gathering information on available instruments, and since 
I assume these folks would like to sell their new system, so why is it that no 
one will provide me with some basic information, like slide capacity, 
capabilities, reagent costs, etc?
Any and all feedback is appreciated.
Jeanine

Sent from Yahoo Mail for iPhone
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Re: [Histonet] IHC and Histogel

[dusko trajkovic via Histonet]

We have used Histogel for years on cell pellets, and have not come across any 
IHC issues.Dusko 

On Thursday, May 10, 2018 7:13 AM, Paula via Histonet 
 wrote:
 

 Hello good morning everyone,

Can someone reply who knows if Histogel causes any interference with IHC
staining?

Thank you in advance,

Paula

Bio-Path Med Group

Fountain Valley, CA

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Re: [Histonet] IHC and Histogel

[John Shelley via Histonet]

I agree with Dusko,

I have used histogel the same way and have also used it for FISH and ISH 
without any issues.

Kind Regards!
 
John J Shelley
Histology Core Manager
Sanford Burnham Prebys Medical Discovery Institute at Lake Nona
6400 Sanger Road    
Orlando, FL 32827    
Tel: (407) 745-2000 Ext.2517
Lab: (407) 745-2119
Fax: (407) 745-2001
email:  jshel...@sbpdiscovery.org   Please note email change


-Original Message-
From: dusko trajkovic via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, May 10, 2018 10:20 AM
To: Paula; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC and Histogel

We have used Histogel for years on cell pellets, and have not come across any 
IHC issues.Dusko 

On Thursday, May 10, 2018 7:13 AM, Paula via Histonet 
<histonet@lists.utsouthwestern.edu> wrote:
 

 Hello good morning everyone,

Can someone reply who knows if Histogel causes any interference with IHC
staining?

Thank you in advance,

Paula

Bio-Path Med Group

Fountain Valley, CA

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Re: [Histonet] IHC validation

[Echague, Caren Ann - MGH via Histonet]

Where do you purchase your Tisssue Micro arrays? I know they are quite 
expensive.
We are currently running validations and using only a handful of positive 
cases. Most of them don't have ten positive and negative cases but as long as 
you can get a statement from the medical director, this should be CAP compliant.
Cae Aguilar, HTL (ASCP)
Histology Supervisor
7079802801

-Original Message-
From: Allan Wang [mailto:all...@biomax.us] 
Sent: Thursday, March 22, 2018 11:02 AM
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC validation

How are labs validating for rarer biomarkers like ALK, ROS1, or MMR loss?
Following the guideline of 10 positive cases may be difficult.
I've seen other companies with control slides just with a few engineered cell 
lines as positive and negative controls. Is that enough for validation alone?

We are creating TMAs for validation of ALK and MMR loss (MLH1, PMS2, MSH6,
MSH2) so I am interested in knowing how many cases are desired.

Allan

Allan Wang
Lab Manager
US Biomax

On Tue, Mar 20, 2018 at 1:54 PM, Terri Braud via Histonet < 
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with 
> the prerequisite number of cases, both positive and negative, and 
> stain your validation all on one slide.  I've done this for years and 
> for 3 different validations of entire IHC platform changes, ranging 
> from 40 to over 100 antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" <plu...@biopath.org>
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a 
> validation for IHC antibodies. We are governed by CLIA, and we would 
> like to know if there is a set number of slides to run for a 
> particular antibody we would like to bring in-house for Validation.  I think 
> CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a 
> certain number of slides, or is it up to us (the laboratory director) 
> to decide how many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
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Re: [Histonet] IHC validation

[Allan Wang via Histonet]

How are labs validating for rarer biomarkers like ALK, ROS1, or MMR loss?
Following the guideline of 10 positive cases may be difficult.
I've seen other companies with control slides just with a few engineered
cell lines as positive and negative controls. Is that enough for validation
alone?

We are creating TMAs for validation of ALK and MMR loss (MLH1, PMS2, MSH6,
MSH2) so I am interested in knowing how many cases are desired.

Allan

Allan Wang
Lab Manager
US Biomax

On Tue, Mar 20, 2018 at 1:54 PM, Terri Braud via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with the
> prerequisite number of cases, both positive and negative, and stain your
> validation all on one slide.  I've done this for years and for 3 different
> validations of entire IHC platform changes, ranging from 40 to over 100
> antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" 
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a validation
> for IHC antibodies. We are governed by CLIA, and we would like to know if
> there is a set number of slides to run for a particular antibody we would
> like to bring in-house for Validation.  I think CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a certain
> number of slides, or is it up to us (the laboratory director) to decide how
> many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
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>
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Re: [Histonet] IHC validation

[Morken, Timothy via Histonet]

I've used hundreds of TMA's from Pantomics and Biochain with great results. 

https://pantomics.com/

https://www.biochain.com/


Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

-Original Message-
From: Dessoye, Michael via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, March 22, 2018 9:00 AM
To: Terri Braud; 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Can anyone recommend a vendor that they've had good luck with for TMA slides?

Michael J. Dessoye, M.S. | Histology/Toxicology/Special Chemistry Supervisor | 
Commonwealth Health Laboratory Services | mjdess...@commonwealthhealth.net | 
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 
570-552-1484

-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com] 
Sent: Tuesday, March 20, 2018 1:54 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" <plu...@biopath.org>
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula




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Re: [Histonet] IHC validation

[Dessoye, Michael via Histonet]

Can anyone recommend a vendor that they've had good luck with for TMA slides?

Michael J. Dessoye, M.S. | Histology/Toxicology/Special Chemistry Supervisor | 
Commonwealth Health Laboratory Services | mjdess...@commonwealthhealth.net | 
575 N. River Street | Wilkes Barre, PA 18764 | Tel: 570-552-1432 | Fax: 
570-552-1484

-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com] 
Sent: Tuesday, March 20, 2018 1:54 PM
To: 'histonet@lists.utsouthwestern.edu'
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" <plu...@biopath.org>
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula




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Re: [Histonet] IHC validation

[Ana Maluenda via Histonet]

Hi Terri,

This is actually a very valid note. This has been in my mind for years, but 
never had the chance to put in action in a research environment (I always try 
to bring to the research field the efficiency and money saving strategies we 
use in diagnostics).

Where do you usually order your unstained slides from?

Much appreciated for the advice.

Kind regards,

Ana

Ana Maluenda
Research Assistant
Atherothrombosis and Vascular Biology Laboratory

Baker Heart and Diabetes Institute
75 Commercial Road, Melbourne VIC 3004
P (03) 8532 1359 E ana.malue...@baker.edu.au W www.baker.edu.au


-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com]
Sent: Wednesday, 21 March 2018 4:54 AM
To: 'histonet@lists.utsouthwestern.edu' <histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] IHC validation

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" <plu...@biopath.org>
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula



Protecting your privacy is important to us. The Baker Heart and Diabetes 
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Re: [Histonet] IHC validation

[Colleen Forster via Histonet]

I agree Terry,

The TMA slide is a very economical and powerful way to validate with
minimal slides needed.

Colleen Forster

On Tue, Mar 20, 2018 at 12:54 PM, Terri Braud via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Just another note:  You can order unstained tissue microarrays with the
> prerequisite number of cases, both positive and negative, and stain your
> validation all on one slide.  I've done this for years and for 3 different
> validations of entire IHC platform changes, ranging from 40 to over 100
> antibodies each time.  Saves time and money.
>
> Terri L. Braud, HT(ASCP)
> Anatomic Pathology Supervisor
> Laboratory
> Holy Redeemer Hospital
> 1648 Huntingdon Pike
> Meadowbrook, PA 19046
> ph: 215-938-3689
> fax: 215-938-3874
> Care, Comfort, and Heal
>
> Message: 2
> Date: Fri, 16 Mar 2018 06:54:30 -0700
> From: "Paula" 
> Subject: [Histonet] Antibody Validation CLIA
>
> Hello,
> We've been discussing about the quantity of slides to run as a validation
> for IHC antibodies. We are governed by CLIA, and we would like to know if
> there is a set number of slides to run for a particular antibody we would
> like to bring in-house for Validation.  I think CAP requires 20 slides..?
> And so we are asking if there is  a requirement with CLIA to run a certain
> number of slides, or is it up to us (the laboratory director) to decide how
> many slides to run for Validation/Verification.
> Thank you in advance
> Paula
>
>
> ___
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> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Re: [Histonet] IHC validation

[Terri Braud via Histonet]

Just another note:  You can order unstained tissue microarrays with the 
prerequisite number of cases, both positive and negative, and stain your 
validation all on one slide.  I've done this for years and for 3 different 
validations of entire IHC platform changes, ranging from 40 to over 100 
antibodies each time.  Saves time and money.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874
Care, Comfort, and Heal

Message: 2
Date: Fri, 16 Mar 2018 06:54:30 -0700
From: "Paula" 
Subject: [Histonet] Antibody Validation CLIA

Hello,
We've been discussing about the quantity of slides to run as a validation for 
IHC antibodies. We are governed by CLIA, and we would like to know if there is 
a set number of slides to run for a particular antibody we would like to bring 
in-house for Validation.  I think CAP requires 20 slides..?
And so we are asking if there is  a requirement with CLIA to run a certain 
number of slides, or is it up to us (the laboratory director) to decide how 
many slides to run for Validation/Verification.
Thank you in advance
Paula


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Re: [Histonet] IHC VALIDATION

[Fulton Regan via Histonet]

Hi Dawn,


I would suggest having a look at this CAP guidance paper, below.  If I can be 
of any assistance, please feel free to contact me. 

1. Patrick, L. F. et al. Principles of analytic validation of 
immunohistochemical assays: Guideline from the College of American Pathologists 
Pathology and Laboratory Quality Center. Arch. Pathol. Lab. Med. 138, 1432–1443 
(2014).


Regan

Regan Fulton, MD/PhD
PhenoPath
551 North 34th Street, Suite 100
Seattle, WA 98103
Phone 206-374-9000   


CEO, Array Science, LLC
475 Gate 5 Road, #102
Sausalito, CA 94965



> On Feb 27, 2018, at 8:34 AM, Olszewski, Dawn via Histonet 
>  wrote:
> 
> Good morning Histo Peeps,
> 
> I was wondering if anyone would be willing to share their protocol on 
> antibody validation for the IHC lab on FFPE tissues.  Do you know if there 
> are specific regulations on validation or does each lab/hospital develop 
> their own methods.  We are no longer under CAP for inspections,  but we are 
> inspected by Joint Commission.
> 
> If you know of specific regulations could you please site them.  We seem to 
> be in disagreement on how this should be done in our lab.
> Thanks for any and all advice in advance.
> 
> Sincerely,
> 
> Dawn Olszewski, HTL(ASCP)QIHC
> ___
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Re: [Histonet] IHC counterstain offline

[Victoria Baker via Histonet]

Curt,

I am not sure how much you will actually save if you factor in the cost of
a person hand counter staining these slides.  I will be the first to say
Ventana is over priced, but off line counter staining brings in an
additional variable and cost.  In the end is it a real savings?  It's an
individual call, but I wouldn't do it.

Vikki

On Oct 12, 2017 4:43 PM, "Curt via Histonet" <
histonet@lists.utsouthwestern.edu> wrote:

> We currently use the benchmark ultra and are, in an effort to save a
> little money, looking at running out counter stain offline. Anyone have any
> suggestions or thoughts they might offer? We currently run our H with
> Richard Allen Hematoxylin 7111. I am curious if that is suitable for use in
> your experience or if you recommend a different Hematoxylin for IHC counter
> stains. How much bluing time, etc.
>
>
>
> Thanks,
>
> Curt
>
>
> CONFIDENTIALITY NOTE: The information transmitted, including attachments,
> is intended only for the person(s) or entity to which it is addressed and
> may contain confidential and/or privileged material. Any review,
> retransmission, dissemination or other use of, or taking of any action in
> reliance upon this information by persons or entities other than the
> intended recipient is prohibited. If you received this in error, please
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Re: [Histonet] IHC

[Dessasau III, Evan via Histonet]

Hi again Ms. Horst,  For some reason I did not see all the replies from Mr. 
Cooper.  But wait before your order the Superfrost slides.  If you are not 
familiar with them they are good as Mr. Cooper says but they can be pricey.  We 
use a great guy at PathSUPPLY, who is cc'd here, that can get you a comparable 
(if not the same) slide at a lower price point.  Also, we use slides packaged 
for PathSupply that are a good price point and excellent quality esp. the 
Bright White slide is comparable to the Superfrost but at a lower price point.  
Also, sorry I would suggest you heat them before you send them and communicate 
with them like someone else said.  I always ask how they want them treated 
before I send the sections.
E-van

-Original Message-
From: Emily Horst via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Thursday, August 10, 2017 6:18 PM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC

Thanks everyone for all your suggestions! You've been very helpful!

On Thu, Aug 10, 2017 at 6:16 PM Cooper, Brian <bcoo...@chla.usc.edu> wrote:

> See if you can get a sample of slides from Thermo Fisher.  We use the
> Superfrost Plus slides, and tissue loss just isn’t one of the problems
> we ever have here.  And like Tim Morken has said in his response,
> ensure there’s no trapped water under the section.
>
>
>
>
> https://www.fishersci.com/shop/products/shandon-colorfrost-plus-slides
> /6776214?searchHijack=true=6776214=RAPID
> CatNo=6776214CS
>
>
>
> Thanks,
>
>
>
> Brian
>
>
>
> *From:* Emily Horst [mailto:mashals...@gmail.com]
> *Sent:* Thursday, August 10, 2017 2:54 PM
>
>
> *To:* Cooper, Brian
> *Subject:* Re: [Histonet] IHC (EXTERNAL EMAIL)
>
>
>
> They are ones that were prepackaged and sent to us from Dianon (Dianon
> is not the lab I'm talking about). I have no idea what/who the slides
> are. I just ordered new charged slides from Mercedes Medical and all
> my invoice says is 45 degree, white. I haven't received them yet so I
> don't have a box with info on it.
>
>
>
> It's a problem we've been having for awhile.
>
>
>
> On Thu, Aug 10, 2017 at 5:49 PM Cooper, Brian <bcoo...@chla.usc.edu>
> wrote:
>
> So I worked at a reference lab for yeears and years and years,
> and I can tell you that not all charged slides are created equally.
> What type of slides are you using?
>
>
>
> Thanks,
>
>
>
> Brian
>
>
>
> *From:* Emily Horst [mailto:mashals...@gmail.com]
> *Sent:* Thursday, August 10, 2017 2:47 PM
> *To:* Cooper, Brian
> *Subject:* Re: [Histonet] IHC (EXTERNAL EMAIL)
>
>
>
> No I am not. I air dry them and then package them up.
>
>
>
> On Thu, Aug 10, 2017 at 5:46 PM Cooper, Brian <bcoo...@chla.usc.edu>
> wrote:
>
> Are you baking the slides before sending them out?  How long, and what
> temperature?
>
> Thanks,
>
> Brian D. Cooper, HT (ASCP)CM | 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 Pager: 213-209-0184
> bcoo...@chla.usc.edu
>
> -Original Message-
> From: Emily Horst via Histonet
> [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Thursday, August 10, 2017 2:42 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] IHC (EXTERNAL EMAIL)
>
> Hi everyone,
>
> We send our IHCs to an outside lab. I send them the unstained slides.
> I cut them at 4 microns and put them on charged slides. We are having
> a lot of issues with the receiving lab saying the tissue is falling
> off. Their supervisor says the sections are too thick (remember 4
> microns) and my pathologist says they are not too thick. Any
> suggestions for resolving this issue with the receiving lab?
>
> Thanks!
> --
> Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
> Kettering, Ohio 937.297.0028
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> CONFIDENTIALITY NOTICE: This e-mail message, including any
> attachments, is for the sole use of the intended recipient(s) and may
> contain confidential or legally privileged information. Any
> unauthorized review, use, disclosure or distribution is prohibited. If
> you are not the intended recipient, please contact the sender by reply
> e-mail and destroy all copies of this original message.
>
> --
>
> Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
> Kettering, Ohio 937.297.0028
>
> CONFIDENTIALITY NOTICE: This e-mail message, including any
> attach

Re: [Histonet] IHC

[Dessasau III, Evan via Histonet]

Hi Emily , we do the same thing.  It could be the slides but I think it has 
more to do  with how you treat the sections after mounting.  The dry time and 
heat time.  I also think it has more to do with technique in the IHC lab.  
Brains, decal, and fatty tissues get more dry time.  All sections get 1 hr. 
heat at 60C .  if they dry overnight 15min heat seems to be fine for us.
E-van

-Original Message-
From: Emily Horst via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Thursday, August 10, 2017 5:42 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC

Hi everyone,

We send our IHCs to an outside lab. I send them the unstained slides. I cut 
them at 4 microns and put them on charged slides. We are having a lot of issues 
with the receiving lab saying the tissue is falling off. Their supervisor says 
the sections are too thick (remember 4 microns) and my pathologist says they 
are not too thick. Any suggestions for resolving this issue with the receiving 
lab?

Thanks!
--
Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio 
Kettering, Ohio 937.297.0028 ___
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This e-mail message (including any attachments) is for the sole use of
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Re: [Histonet] IHC

[Emily Horst via Histonet]

Thanks everyone for all your suggestions! You've been very helpful!

On Thu, Aug 10, 2017 at 6:16 PM Cooper, Brian <bcoo...@chla.usc.edu> wrote:

> See if you can get a sample of slides from Thermo Fisher.  We use the
> Superfrost Plus slides, and tissue loss just isn’t one of the problems we
> ever have here.  And like Tim Morken has said in his response, ensure
> there’s no trapped water under the section.
>
>
>
>
> https://www.fishersci.com/shop/products/shandon-colorfrost-plus-slides/6776214?searchHijack=true=6776214=RAPID=6776214CS
>
>
>
> Thanks,
>
>
>
> Brian
>
>
>
> *From:* Emily Horst [mailto:mashals...@gmail.com]
> *Sent:* Thursday, August 10, 2017 2:54 PM
>
>
> *To:* Cooper, Brian
> *Subject:* Re: [Histonet] IHC (EXTERNAL EMAIL)
>
>
>
> They are ones that were prepackaged and sent to us from Dianon (Dianon is
> not the lab I'm talking about). I have no idea what/who the slides are. I
> just ordered new charged slides from Mercedes Medical and all my invoice
> says is 45 degree, white. I haven't received them yet so I don't have a box
> with info on it.
>
>
>
> It's a problem we've been having for awhile.
>
>
>
> On Thu, Aug 10, 2017 at 5:49 PM Cooper, Brian <bcoo...@chla.usc.edu>
> wrote:
>
> So I worked at a reference lab for yeears and years and years, and
> I can tell you that not all charged slides are created equally.  What type
> of slides are you using?
>
>
>
> Thanks,
>
>
>
> Brian
>
>
>
> *From:* Emily Horst [mailto:mashals...@gmail.com]
> *Sent:* Thursday, August 10, 2017 2:47 PM
> *To:* Cooper, Brian
> *Subject:* Re: [Histonet] IHC (EXTERNAL EMAIL)
>
>
>
> No I am not. I air dry them and then package them up.
>
>
>
> On Thu, Aug 10, 2017 at 5:46 PM Cooper, Brian <bcoo...@chla.usc.edu>
> wrote:
>
> Are you baking the slides before sending them out?  How long, and what
> temperature?
>
> Thanks,
>
> Brian D. Cooper, HT (ASCP)CM | 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 Pager: 213-209-0184
> bcoo...@chla.usc.edu
>
> -Original Message-
> From: Emily Horst via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Thursday, August 10, 2017 2:42 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] IHC (EXTERNAL EMAIL)
>
> Hi everyone,
>
> We send our IHCs to an outside lab. I send them the unstained slides. I
> cut them at 4 microns and put them on charged slides. We are having a lot
> of issues with the receiving lab saying the tissue is falling off. Their
> supervisor says the sections are too thick (remember 4 microns) and my
> pathologist says they are not too thick. Any suggestions for resolving this
> issue with the receiving lab?
>
> Thanks!
> --
> Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
> Kettering, Ohio 937.297.0028 ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> CONFIDENTIALITY NOTICE: This e-mail message, including any attachments,
> is for the sole use of the intended recipient(s) and may contain
> confidential
> or legally privileged information. Any unauthorized review, use, disclosure
> or distribution is prohibited. If you are not the intended recipient,
> please
> contact the sender by reply e-mail and destroy all copies of this original
> message.
>
> --
>
> Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
> Kettering, Ohio 937.297.0028
>
> CONFIDENTIALITY NOTICE: This e-mail message, including any attachments,
> is for the sole use of the intended recipient(s) and may contain
> confidential
> or legally privileged information. Any unauthorized review, use, disclosure
> or distribution is prohibited. If you are not the intended recipient,
> please
> contact the sender by reply e-mail and destroy all copies of this original
> message.
>
> --
>
> Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
> Kettering, Ohio 937.297.0028
>
> CONFIDENTIALITY NOTICE: This e-mail message, including any attachments,
> is for the sole use of the intended recipient(s) and may contain
> confidential
> or legally privileged information. Any unauthorized review, use, disclosure
> or distribution is prohibited. If you are not the intended recipient,
> please
> contact the sender by reply e-mail and destroy all copies of this original
> message.
>
-- 
Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio
Kettering, Ohio 937.297.0028
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Re: [Histonet] IHC

[Morken, Timothy via Histonet]

Emily, ? Are you ensuring excess water is not under the section that would 
prevent adherence to the slide? Are you using slides recommended by the outside 
lab? Do they recommend a procedure for drying the slides before sending?
Just a starting point...

Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center

-Original Message-
From: Emily Horst via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Thursday, August 10, 2017 2:42 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC

Hi everyone,

We send our IHCs to an outside lab. I send them the unstained slides. I cut 
them at 4 microns and put them on charged slides. We are having a lot of issues 
with the receiving lab saying the tissue is falling off. Their supervisor says 
the sections are too thick (remember 4 microns) and my pathologist says they 
are not too thick. Any suggestions for resolving this issue with the receiving 
lab?

Thanks!
--
Emily Horst, HT (ASCP) Office manager Interventional Pathology of Ohio 
Kettering, Ohio 937.297.0028 ___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
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Re: [Histonet] IHC questions

[Tony Henwood (SCHN) via Histonet]

Hi Vicki,



We recently did a study that we published in our local journal (Histograph June 
2017) that migh be of use:



Controlled Section Baking for Immunohistochemistry
Tony Henwood, Principal Scientist, Histopathology, The Children's Hospital at 
Westmead

One source of poor immunostaining is overheating of tissue and sections. 
Several authors have reported that heated slide drying adversely affects 
sensitivity in immunohistochemistry (1). Therefore, some have advocated the use 
of lower temperature drying using adhesive-coated slides to improve the 
sensitivity of the test (2-5).

In one study (1), half the antigens were adversely affected by section drying 
at 80oC including 5D3, CMV, S100, HMB45 and CEA. Oates (4) used antisera to 
epithelial membrane antigen from three different companies and found that for 
slides dried at 58"C, staining was often paler than slides dried at room 
temperature or at 37°C.

Low heat attachment of sections to slides can cause several issues including 
inadequate attachment of the tissue sections so that tissue sections may be 
lost during antigen recovery and/or immunostaining, the inability of some 
paraffins to melt well at 58oC, and the requirement of more than 1 hr before an 
immunohistochemical procedure may be started. It has been recommended that the 
most efficient protocol for mounting tissue sections to microscopic slides 
would be to attach the tissues overnight before applying the 
immunohistochemical procedure at a temperature at which all tissue mounting 
paraffins should melt (e.g., 65oC) (6). It should be remembered that a 
significant dewaxing of sections occurs when slides are heated a few degrees 
above the melting point of the wax.
Laboratory ovens seem to be variable in their ability to maintain a constant 
temperature with the implication that it is possible to either over-cook 
sections thus adversely affecting antigens or under-heat them, possibly 
compromising subsequent de-waxing. There is also the human element. How often 
are slides removed from the oven at the required time?

The modern automatic immunostainers have excellent on-board slide heating in 
order to achieve reproducible, accurate antigen retrieval. This feature also 
allows controlled "baking" of sections and being able to programme a set time, 
removes the possibility of human error. At the Children's Hospital, the Bond 3 
is used for automated immunohistochemistry. A study was designed to assess the 
usefulness of on-board baking in routine immunohistochemistry.

Control sections were immunostained for several antigens (see table) using the 
Bond 3 on-board baking and dewax facility. Freshly cut sections were dried at 
37oC for 5 minutes to remove excess water. Slides were then loaded onto the 
Bond and the baking procedure used was 35 minutes at 63oC. Stained controls 
were compared with control slides stained prior to the instigation of the 
on-board bake procedure. The historic procedure involved heating sections at 
63-65oC for 35 minutes in a large fan-forced dry-air oven (7).

BCL-2

Mum-1

CD31

BCL-6

Calretinin

SATB2

BOB-1

S100

CyclinD1

CD20

ALK-1

MPO

CD21

Ki67

INI-1

CD3

Synaptophsin

BRG-1

HMB-45

Chromogranin

Inhibin

Melan A

Myogenin

Desmin


The results showed that there was no difference between controls stained with 
the historic compared to the on-board baking procedure except for BCL-6 which 
the new procedure gave stronger staining. (see figure).

In conclusion, we expect that on-board baking of sections should allow 
laboratories to have better control over the pre-analytical variables that can 
adversely affect the immunohistochemistry staining.

References

1.   Henwood, A. F. (2005). Effect of slide drying at 80oC on 
immunohistochemistry. Journal of Histotechnology, 28(1), 45-46.

2.   Wakins, J., Kellock, D., Gillet, C., Egan, M., Pontin, J. E., Millis, 
R. R., & Levinson, D. A. (1990). Enhancement of immunostaining. Histopathology, 
17(2), 185-185.

3.   Dodson, A., Davies, E., & Waring, J. (1991). APTES, a section adhesive 
for immunocytochemistry; and experiences of slide drying at room temperature. 
Histopathology, 19(5), 484-485.

4.   Oates J. (1993) The effect of temperature on immunostaining. Br J 
Biomed Sci 50: 157-158,

5.   Williams, J. H., Mepham, B. L., & Wright, D. H. (1997). Tissue 
preparation for immunocytochemistry. Journal of clinical pathology, 50(5), 
422-428.

6.   Jones, W. T., Stockard, C. R., & Grizzle, W. E. (2001). Effects of 
time and temperature during attachment of sections to microscope slides on 
immunohistochemical detection of antigens. Biotechnic & Histochemistry, 76(2), 
55-58.

7.   Henwood, A. F. (2012). The application of heated detergent dewaxing 
and rehydration to immunohistochemistry. Biotechnic & Histochemistry, 87(1), 
46-50.





Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)

Principal Scientist, the Children's Hospital at 

Re: [Histonet] IHC sections adherence

[Kari Simeone via Histonet]

Hi Greg, I use BONDS and Apex slides as well. What we do with tissue that 
washes (usually heavy tumor and some harsher stains like CD34) is leave the 
tissue unbaked at room temp for 12 hours and then bake at 60 degrees for 30 
minutes. I don't know if you are including your control tissue on the same 
slide (we do) but we also cut a separate slide with just patient sections and 
that seems to help with extra paraffin runoff from the control tissue 
underneath the patient tissue. We've been doing this successfully for years.
I hope this is helpful, I totally understand your frustration! 

Kari M Simeone

Histology/Immunohistochemistry Specialist Supervisor



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Subject: Histonet Digest, Vol 164, Issue 9

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Today's Topics:

   1. Re: Section adherence issues (IHC) (Walter Benton)
   2. Permanent/Full Time Histotech Job in Panama City, FL
  (Melissa Owens)
   3. New Lab Related Blog Post (Lester Raff MD)


--

Message: 1
Date: Wed, 12 Jul 2017 17:37:38 +
From: Walter Benton 
To: "Hujet, Matthew" 
, Greg Dobbin


Cc: "kgevele...@ihis.org" ,
"histonet@lists.utsouthwestern.edu"

Subject: Re: [Histonet] Section adherence issues (IHC)
Message-ID: <816e042155824c4193f79c50c1461...@mail01.gcu-MD.local>
Content-Type: text/plain; charset="us-ascii"

Thanks. This is helpful information.

-Original Message-
From: Hujet, Matthew [mailto:matthew.hu...@ssmhealth.com]
Sent: Wednesday, July 12, 2017 1:35 PM
To: Walter Benton ; Greg Dobbin 

Cc: kgevele...@ihis.org; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Section adherence issues (IHC)

We have had really good luck with both the TOMO and Millennia 2000 adhesion 
slides. We initially tried using the TOMO slides for everything (IHC, special 
stains, and H's), but found that there was too much background staining on 
the specials and H's. For IHC, the TOMO slides were great. We then switched 
to the Millennia 2000 for everything, and they have been working really well 
across the board. With regards to just IHC, I personally feel that the TOMO 
slides are better, but if you only want one type of glass in your lab, the 
Millennia 2000 are the way to go.

Matthew Hujet
Histology Technical Specialist
SSM Health St. Mary's Hospital - Madison
700 S. Park St.
Madison, WI 53715

Please note my new email address: matthew.hu...@ssmhealth.com



-Original Message-
From: Walter Benton [mailto:wben...@cua.md]
Sent: Wednesday, July 12, 2017 10:34 AM
To: Greg Dobbin 

Cc: kgevele...@ihis.org; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Section adherence issues (IHC)

Greg,

We are running into a similar issue on the Biocare Nemesis platform and the 
support there suggest humidity to be the issue. Our sections stay adhered to 
the slides through depar, antigen retrieval, but 

Re: [Histonet] IHC for secreted proteins and cytokines in frozen sections - Pre-fix or post-fix?

[Morken, Timothy via Histonet]

Ana, you should fix before sectioning. Chris Van der Loos did an excellent 
study on loss of cytokines in frozen sections vs whole cells. See his paper:

Immunohistochemical Detection of Interferon-y, Fake or Fact? CM Van der Loos, 
et.al., J Histochem Cytochem, 49(6):699-709, 2001.  Www.jhc.org



Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center




-Original Message-
From: Ana Maluenda via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Friday, June 02, 2017 1:01 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC for secreted proteins and cytokines in frozen sections 
- Pre-fix or post-fix?

Hi everyone,

Just wondering what is people's opinion and protocols for IHC in mouse frozen 
sections targeting secreted proteins and cytokines. I see lots of places using 
fresh frozen sections/snap-freeze and cold acetone or methanol/ethanol 
post-fixation. Is this an issue when it comes to diffusion of such proteins in 
the tissue, since they are not well localized or linked to cellular structures? 
Would it be better to pre-fix (either immersion in 4%PFA or infusion with 
fixative) than post-fix?

Any thoughts would be much appreciated.

Kind regards,

Ana

Ana Maluenda
Research Assistant


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Re: [Histonet] IHC testing for Parvovirus (Human)

[Tony Henwood (SCHN) via Histonet]

Hi Richard,
We use NCL-PARVO from Leica

Details as follows:

ANTIGEN NAME:Parvovirus B19
OTHER NAMES: 

Clone:  R92F6
Isotype:IgG1  


DESCRIPTION:  Mouse monoclonal to native parvovirus B19 purified from human 
plasma. Human parvovirus B19 (VP1 and VP2 capsid proteins).

STORAGE CONDITIONS:4-8oC undiluted

SUPPLIER:Leica  NCL-PARVO
PROCEDURE:

Methodology:HRP-POLYMER Working Dilution:   Bond: 1/100
Special Conditions:  Citrate HIER required (ER1) 
CONTROL TISSUE: Placenta containing Parvovirus   

Inbuilt Controls 
CLINICAL SIGNIFICANCE:
Diagnosis of parvovirus B19 infection in formalin-fixed, paraffin embedded 
tissue can be made through the identification of the characteristic brick-red 
intranuclear inclusions in normoblasts within the fetal circulation in 
histological sections stained with HE. It has been found that 
immunohistochemistry was the best detection method. It is highly specific and 
sensitive, preserves the morphology and reveals a larger number of positive 
cells than does HE with the advantage of showing cytoplasmic and nuclear 
positivity, making it more reliable.
REFERENCES:
Li, J. J., Henwood, T., Van Hal, S., & Charlton, A. (2015). Parvovirus 
infection: an immunohistochemical study using fetal and placental tissue. 
Pediatric and Developmental Pathology, 18(1), 30-39.
Quemelo PRV, Lima DM, Fonseca BAL, Peres LC (2007) "Detection of parvovirus B19 
infection in formalin-fixed and paraffin-embedded placenta and fetal tissues" 
Rev. Inst. Med. trop. S. Paulo, 49(2): 103-107,

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-Original Message-
From: Cartun, Richard via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Wednesday, 24 May 2017 4:33 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC testing for Parvovirus (Human)

If you do this test, where do you get your antibody?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic 
Proteomics Laboratory Director, Biospecimen Collection Programs Assistant 
Director, Anatomic Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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Re: [Histonet] IHC billing on archived case

[Weems, Joyce K. via Histonet]

However, if it is a Medicare Patient - it would now be an archived case and 
would be treated as non-hospital. Medicare should pay. j

Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
770-380-8099 Cell
joyce.we...@emoryhealthcare.org



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5665 Peachtree Dunwoody Road
Atlanta, GA 30342

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review, use, disclosure, or distribution is prohibited. If you are not the 
intended recipient, please delete this message, and reply to the sender 
regarding the error in a separate email.

-Original Message-
From: Rene J Buesa via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Wednesday, March 22, 2017 12:28 PM
To: Haines, Beth <bhai...@caperegional.com>; 
'histonet@lists.utsouthwestern.edu' <histonet@lists.utsouthwestern.edu>
Subject: Re: [Histonet] IHC billing on archived case

You should treat it as a new request.René

On Wednesday, March 22, 2017 11:25 AM, "Haines, Beth via Histonet" 
<histonet@lists.utsouthwestern.edu> wrote:


 Hello all,
After some discussion on IHC billing, I have been asked to verify accepted 
billing practice for the following situation:
An IHC request for 2 single antibody stains has been made by outpatient 
oncology practice on a case (same block) that was completed nearly a year ago. 
6 IHCs were done initially (billed as 88342 & 88341 x 5). How should the new 
request be billed? Should a new visit/encounter be created and 88342 & 88341 be 
billed for this encounter?
Thanks in advance for the guidance.


Beth Haines BS, HTL (ASCP)
bhai...@caperegional.com<mailto:bhai...@caperegional.com>
Histology Supervisor
Cape Regional Medical Center

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Re: [Histonet] IHC billing on archived case

[Rene J Buesa via Histonet]

You should treat it as a new request.René 

On Wednesday, March 22, 2017 11:25 AM, "Haines, Beth via Histonet" 
 wrote:
 

 Hello all,
After some discussion on IHC billing, I have been asked to verify accepted 
billing practice for the following situation:
An IHC request for 2 single antibody stains has been made by outpatient 
oncology practice on a case (same block) that was completed nearly a year ago. 
6 IHCs were done initially (billed as 88342 & 88341 x 5). How should the new 
request be billed? Should a new visit/encounter be created and 88342 & 88341 be 
billed for this encounter?
Thanks in advance for the guidance.


Beth Haines BS, HTL (ASCP)
bhai...@caperegional.com
Histology Supervisor
Cape Regional Medical Center

Confidentiality Notice: This e-mail message, including any attachments, from 
Cape
Regional Health System contains information which is CONFIDENTIAL AND/OR LEGALLY
PRIVILEGED.The information is intended only for the use of the individual named
above and may not be disseminated to any other party without Cape Regional 
Health
System's written permission. If you are not the intended recipient, or the
employee or agent responsible for delivering the message to the intended
recipient, you are hereby notified that any dissemination, disclosure,
distribution, copying or taking of any action in reliance on the contents of 
this
e-mailed information is strictly prohibited.  If you have received this e-mail 
in
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the return of these documents to us without cost to you.


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Re: [Histonet] IHC on alcoholic fixed cytology smears

[Rene J Buesa via Histonet]

M best advise is contacting Dako and obtain from them the IHC manual, which 
covers essentially everything you need to know on this subject.René 

On Saturday, March 4, 2017 2:51 PM, Lynette Pavelich via Histonet 
 wrote:
 

 Hello,
I will soon be starting the process of validating many cytoplasmic,
membranous, and nuclear antibodies on alcoholic fixed cytology smears. They
will be performed using the Bond III.

In doing research on the subject, I am finding many different variables,
and it is starting to be confusing. Is anyone willing to share their
processes, or can suggest good reading material for IHC with alcohol fixed
cytology smears? Hints, tips, tried and true methods would be greatly
appreciated.

Thank you,
Lynette Pavelich, HT(ASCP)
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Re: [Histonet] IHC Personel

[Terri Braud via Histonet]

This should help...the IHC "test" is the pathologists' INTERPRETATION of IHC 
stain.  The stain procedure, validation of protocols and controls, and lot to 
lot validation must be signed off by a pathologist.
 Any tech that has demonstrated competency in performing the procedure, can 
perform IHC staining. Any tech can also perform an antibody work-up, provided 
the "results" are signed off by a pathologist.
Easy-peasy

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Laboratory
Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
ph: 215-938-3689
fax: 215-938-3874

-Original Message-

   4. Personel (Jesus Ellin)
--
Message: 4
Date: Tue, 22 Nov 2016 17:36:20 +
From: Jesus Ellin 
Subject: [Histonet] Personel
So I know I am going to open Pandoras box,, but have people been paying 
attention to the Personal requirements from CAP.

I called the CAP and asked them about the criteria concerning Moderate or High 
complexity testing, after discussing with them the situations,   IF you have a 
tech that is Licensed and Also has a QIHC, but does not minimum requirement 
Defined by CLIA in education ,, they CAN NOT do any QA/OC of IHC and antibody 
work up,, as IHC is defined as High complexity testing.

I also asked about the test systems.  The grandfather clause is only good for 
test systems that occurred for those time periods.  For instance if CLIA 
defined the test system after those dates of 1997,, then they are not included 
and the person cannot perform test and technology created after those dates, 
since the testing was not in place during the grandfather clause time.  In a 
nut shell meaning if the IHC staining and antibody was developed after those 
dates,, you are not covered by the grandfather clause to do the testing ,, can 
some help clear this up,,

So any help on this matter will do




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Re: [Histonet] IHC positive controls

[Morken, Timothy via Histonet]

Cynthia, best practice is to keep in the block and cut as needed. Second is to 
cut and dry at room temp and not melt, then store. Don't cut, melt and store. 
The issue is oxidation. Keeping in paraffin prevents oxidation. You'll protect 
the antigens better if they  are fully isolated from air, which paraffin does, 
but once cut, they are exposed. That happens in the fridge as well and I'm not 
sure refrigeration helps much. I'm guessing that you don't do a lot of these, 
so you don't want to cut a lot of slide and have them sitting around for months 
waiting for use. 


Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center




-Original Message-
From: Cindy Bulmer via Histonet [mailto:histonet@lists.utsouthwestern.edu] 
Sent: Monday, May 02, 2016 10:43 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC positive controls


Hello Histoland,
 
I need some advice, I have a PT block that is positive with Spirochetes.
What would be the best way to use this block as a positive control?
 
1)  Cut (serial sections, stain the last slide for bugs) and oven time (60) for 
1 hr. 
 then put slides in refrigerator for future use.
2)  Cut (serial sections, stain the last slide for bugs) NO oven time  and put 
slides
 directly in refrigerator for future use.
3)  Cut "fresh" every time they order the Ab.
 
Thank you,
Cindy


Cynthia Bulmer HT(ASCP),QIHC
IHC Supervisor, CTPL
Waco, TX




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Re: [Histonet] IHC with H & E staining

[Amos Brooks via Histonet]

Hi,
I usually try to avoid eosin as a counterstain for a DAB labeled slide
because the red/pink of the eosin can obscure the rusty brown of DAB. If
you really want to use it though I would suggest a *really* light eosin,
perhaps even just a few milliliters in the 95% ETOH as you are dehydrating
it. The RBCs and eosinophils will pick it up quickly and the stain should
not overwhelm the DAB. You could also darken the DAB with Copper sulfate.

Cheers,
Amos

On Wed, Jan 6, 2016 at 1:00 PM, 
wrote:

> Message: 6
> Date: Wed, 6 Jan 2016 11:26:35 +
> From: Marcus Green 
> To: "histonet@lists.utsouthwestern.edu"
> 
> Subject: [Histonet] IHC with H & E staining
> Message-ID:
> <669331a24821fe4491665e8dfd47cf22c25...@mbx06.ad.oak.ox.ac.uk>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Dear Users,
>
>
>
> Happy New year - I hope this finds everybody well!
>
>
>
> I was asked a very simple question yesterday - why don't you do H
> counterstaining on DAB stained samples. The question came about as we're
> looking at CD31 staining for blood-vessels and some look ruptured (we're
> keen to see red blood cells).
>
>
>
> Instead of doing a consecutive cuts, a CD31 stain (Haem counterstain) on
> one slide and H on another, the question was asked why not do the Eosin
> stain as part of the counterstain
>
>
>
> I've never seen it done in the literature or in the clinic, and I've never
> asked why it's not done. Any assistance or advice would be greatly welcome
> - and my sincerest apologies if this is a very basic question?
>
>
>
> Thanks in advance for your time,
>
> kind regards,
>
>
>
> Marcus,
>
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Re: [Histonet] IHC with H & E staining

[Elizabeth Chlipala via Histonet]

Marcus

You can stain with H after DAB if you think it would help your how you 
analyze your slides.  DAB with the addition of a special stain has been 
published and for us depending upon the project that we are working on we may 
choose to use a different counterstain after an IHC that has DAB as an 
chromogen.  For example we have worked with macrophage markers counterstained 
with Prussian blue, another example is brain IHC markers counterstained Cresyl 
ect Violet rather than hematoxylin.  DAB is extremely stable and therefore many 
special stains or different counterstains can be used.  

If you are running an image analysis algorithm most canned algorithms are set 
up to function with a hematoxylin counterstain - you would need evaluate if a 
different counterstain would decrease the accuracy of your algorithm but you 
may have the ability to design an algorithm with the counterstain of your 
choice.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
l...@premierlab.com

Ship to address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

From: Marcus Green via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Wednesday, January 06, 2016 4:26 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC with H & E staining

Dear Users,



Happy New year - I hope this finds everybody well!



I was asked a very simple question yesterday - why don't you do H 
counterstaining on DAB stained samples. The question came about as we're 
looking at CD31 staining for blood-vessels and some look ruptured (we're keen 
to see red blood cells).



Instead of doing a consecutive cuts, a CD31 stain (Haem counterstain) on one 
slide and H on another, the question was asked why not do the Eosin stain as 
part of the counterstain



I've never seen it done in the literature or in the clinic, and I've never 
asked why it's not done. Any assistance or advice would be greatly welcome - 
and my sincerest apologies if this is a very basic question?



Thanks in advance for your time,

kind regards,



Marcus,


Department of Oncology - University of Oxford,
Old Road Campus Research Building,
Roosevelt Drive,
Oxford,
OX3 7DQ


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Re: [Histonet] IHC on old slides

[Rene J Buesa via Histonet]

The only thing you can do is to prolong the Ab staining time but, in order to 
avoid excessive background, dilute it. You will have to find an adequate 
balance between a more diluted Ab with a weaker spitope signal and a  prolonged 
incubation time and there is no "magic formula" for obtaining it. You will have 
to make tests.Antigen in stored slides oxidizes producing a weaker signal.René  


 On Friday, November 13, 2015 10:51 AM, "Stoll, Kathryn via Histonet" 
 wrote:
   

 Hi Everyone,

I have been doing some IHC staining on slides that are 20 years old or more.  
Depending on the antibody the staining is pale in comparison to my control 
slide.  Does anyone have experience with staining older slides?  Do you have 
any tips to share?  The slide storage conditions are not something I can 
control.  I am under the assumption that they were stored at room temperature.
Thank you in advance.

Kathryn Stoll

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Re: [Histonet] IHC on old slides

[Hobbs, Carl via Histonet]

Amplification is the only way.

Possibly, tyramide amplification will help.

It will cost you to buy a kit...sure, make your own butyou have to weigh up 
pros/cons


Sureyou will have to play with variables.

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Re: [Histonet] IHC hand staining

[Hughes, Anna [JRDUS] via Histonet]

Hi!
I have done a lot of manual staining in the pasta simple protocol is below: 
 Any of the Dako kits are wonderful...as are the Biocare

1. Depar slides as usual 
2.Place slides in DPBS (no mag or calcium) 2x 5 minutes
3. Antigen retrieval if necessary - will have to look at the insert for your 
primary antibody.
4. Block Peroxidases for 10 minutes with 3% H2O2
5. Wash using DPBS 2x 5minutes
6. Protein block - 30 minutes using 5% normal serum of the host of your 
secondary antibody
7.Wash using DPBS 2x 5 minutes
8. Place slides in your primary antibody (I usually use 200-300ul/slide 
depending on tissue size and cover with a small piece of parafilm).  Incubate 
in a humidity box at RT on the bench for 1 hour
9. Wash using DPBS 2x5 minutes
10. Place slides in a biotinylated secondary antibody (incubate same as primary 
above)- for 20 minutes
11. Wash using DPBS 2x5 mintues
12. Use a DAB detection kit such as the DAKO Envision kit - follow their 
directions in the kit. (These come as mouse,rabbit, rat.. so if you buy the 
kit as a whole, it comes with the biotinylated secondary (goat host) - already 
diluted and ready to use)
13. Wash 2x dH2O
14. Counterstain with Hematoxylin for 5 minutes
15. Wash with dH2O 2x 5 minutes
16. Blue slides  -this will depend on your hematoxylin
17. Wash with dH2O 2x5minutes
18. Dehydrate through alcohols to xylene and coverslip.

Hope this is helpful!
Anna Hughes


-Original Message-
From: Jennifer MacDonald [mailto:jmacdon...@mtsac.edu] 
Sent: Thursday, November 05, 2015 6:20 PM
To: Elaine allison Hoffman
Cc: Histonet List
Subject: Re: [Histonet] IHC hand staining

In one of our classes the students do IHC staining by hand.  They perform 
multiple procedures over the semester.  We currently use Biocare reagents. 
 The polymer based detection systems are simple to use.  The data sheets that 
accompany the antibodies and detection reagents are really helpful with 
protocols. 



From:   Elaine allison Hoffman via Histonet 
<histonet@lists.utsouthwestern.edu>
To: Histonet List <histonet@lists.utsouthwestern.edu>
Date:   11/05/2015 08:08 AM
Subject:[Histonet] IHC hand staining



Greetings Histonetters,
I need some information on "hand" immuno-staining.  We are a small GI lab doing 
Giemsa stains on H. Pylori right now, but the Lab Director wants me to look 
into IHC hand staining for H.Pylori.  Anybody out there doing hand staining and 
willing to give me a protocol to try? Or maybe point me in the right direction 
to who could help me with that.  Any reliable IHC staining kits recommended?  
Thanks in advance for the help
Elaine Hoffman
The Gastroenterology Clinic & Endoscopy Center, IncGI Pathology 
Laboratory1622 E Market StreetWarren, Ohio   44483

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Re: [Histonet] IHC hand staining

[Jennifer MacDonald via Histonet]

In one of our classes the students do IHC staining by hand.  They perform 
multiple procedures over the semester.  We currently use Biocare reagents. 
 The polymer based detection systems are simple to use.  The data sheets 
that accompany the antibodies and detection reagents are really helpful 
with protocols. 



From:   Elaine allison Hoffman via Histonet 

To: Histonet List 
Date:   11/05/2015 08:08 AM
Subject:[Histonet] IHC hand staining



Greetings Histonetters,
I need some information on "hand" immuno-staining.  We are a small GI lab 
doing Giemsa stains on H. Pylori right now, but the Lab Director wants me 
to look into IHC hand staining for H.Pylori.  Anybody out there doing hand 
staining and willing to give me a protocol to try? Or maybe point me in 
the right direction to who could help me with that.  Any reliable IHC 
staining kits recommended?  Thanks in advance for the help
Elaine Hoffman
The Gastroenterology Clinic & Endoscopy Center, IncGI Pathology 
Laboratory1622 E Market StreetWarren, Ohio   44483

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Re: [Histonet] IHC hand staining

[Carlos Defeo via Histonet]

IHC staining for H.Pylori could be done using monoclonals from 
CellMarque or Biocare, with a simple protocol reccommended by the 
vendor,

My regards,
Carlos
Histotochnologist

-- Mensaje original --
De: "Elaine allison Hoffman via Histonet" 


Para: "Histonet List" 
Enviado: 05/11/2015 13:05:38
Asunto: [Histonet] IHC hand staining


Greetings Histonetters,
I need some information on "hand" immuno-staining.  We are a small GI 
lab doing Giemsa stains on H. Pylori right now, but the Lab Director 
wants me to look into IHC hand staining for H.Pylori.  Anybody out 
there doing hand staining and willing to give me a protocol to try? Or 
maybe point me in the right direction to who could help me with that.  
Any reliable IHC staining kits recommended?  Thanks in advance for the 
help

Elaine Hoffman
The Gastroenterology Clinic & Endoscopy Center, IncGI Pathology 
Laboratory1622 E Market StreetWarren, Ohio   44483


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Re: [Histonet] IHC hand staining

[Loralei Dewe via Histonet]

I learned how to do all my IHC by hand from processing and embedding the
tissue to doing the cutting (manual microtome) and subsequent staining.  It
has really helped me in evaluating assays and in troubleshooting customers
problems!

Loralei Dewe
Chrysalis Innovations
On Nov 5, 2015 3:39 PM, "Carlos Defeo via Histonet" <
histonet@lists.utsouthwestern.edu> wrote:

> IHC staining for H.Pylori could be done using monoclonals from CellMarque
> or Biocare, with a simple protocol reccommended by the vendor,
> My regards,
> Carlos
> Histotochnologist
>
> -- Mensaje original --
> De: "Elaine allison Hoffman via Histonet" <
> histonet@lists.utsouthwestern.edu>
> Para: "Histonet List" 
> Enviado: 05/11/2015 13:05:38
> Asunto: [Histonet] IHC hand staining
>
> Greetings Histonetters,
>> I need some information on "hand" immuno-staining.  We are a small GI lab
>> doing Giemsa stains on H. Pylori right now, but the Lab Director wants me
>> to look into IHC hand staining for H.Pylori.  Anybody out there doing hand
>> staining and willing to give me a protocol to try? Or maybe point me in the
>> right direction to who could help me with that.  Any reliable IHC staining
>> kits recommended?  Thanks in advance for the help
>> Elaine Hoffman
>> The Gastroenterology Clinic & Endoscopy Center, IncGI Pathology
>> Laboratory1622 E Market StreetWarren, Ohio   44483
>>
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>>
>
>
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Re: [Histonet] IHC Weekend Coverage

[Davis, Cassie via Histonet]

No IHC/HC/HE weekend coverage. If there is a Stat the supervisor comes in.

Cassandra Davis
Histology Technician
Anatomical Pathology Laboratory
Saint Francis Healthcare
701 N. Clayton Street
Wilmington,DE 19805
Office:  302-575-8095
Email:  cda...@che-east.orgmailto:n...@che-east.org
www.saintfrancishealthcare.org


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Re: [Histonet] IHC Weekend Coverage

[Noelle Linke via Histonet]

Nope

Thank you,
Noëlle

Noëlle Linke, MS, HTL(ASCP) QIHC
Manager, Anatomic Pathology
Pacific Diagnostic Laboratories
nli...@sbch.org
Phone: (805) 324-9814
Fax: (805) 696-6433

-Original Message-
From: Cartun, Richard via Histonet [mailto:histonet@lists.utsouthwestern.edu]
Sent: Sunday, August 23, 2015 7:29 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Weekend Coverage

How many of you working in hospital-based pathology laboratories run IHC on 
weekends?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic 
Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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Re: [Histonet] IHC weekend coverage

[Blake Taylor via Histonet]

We have Saturday lab coverage which consists of a single histotech and only 
inpatient OR samples are cut.  We do not perform IHC or specials on the weekend.

Blake Taylor

-Original Message-
From: histonet-requ...@lists.utsouthwestern.edu 
[mailto:histonet-requ...@lists.utsouthwestern.edu] 
Sent: Sunday, August 23, 2015 1:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 141, Issue 21

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Contents of Histonet digest...


Today's Topics:

   1. F480 antibody (Laurie Finch)
   2. IHC Weekend Coverage (Cartun, Richard)
   3. Re: IHC Weekend Coverage (Victoria Baker)


--

Message: 1
Date: Sat, 22 Aug 2015 18:42:42 -0500
From: Laurie Finch riepo...@gmail.com
To: litepath2...@yahoo.com, histonet@lists.utsouthwestern.edu
Subject: [Histonet] F480 antibody
Message-ID:
CAMjAh1RLWakVPXN8=4j7eooeqb41mm7s-kdncd1edqoqvod...@mail.gmail.com
Content-Type: text/plain; charset=UTF-8

Abserotec has a great f4/80 antibody.

Laurie


--

Message: 2
Date: Sun, 23 Aug 2015 14:28:46 +
From: Cartun, Richard richard.car...@hhchealth.org
To: histonet@lists.utsouthwestern.edu
histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC Weekend Coverage
Message-ID:
9215bd4b0ba1b44d962a71c758b68d2e6b037...@hhcexchmb03.hhcsystem.org
Content-Type: text/plain; charset=us-ascii

How many of you working in hospital-based pathology laboratories run IHC on 
weekends?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic 
Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax


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--

Message: 3
Date: Sun, 23 Aug 2015 10:54:32 -0400
From: Victoria Baker bakevicto...@gmail.com
To: Cartun, Richard richard.car...@hhchealth.org
Cc: histonet netserver histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC Weekend Coverage
Message-ID:
calyzrmqtzxjczimoc6noy-mm13syrkbbpmy6gbgafpb0lp9...@mail.gmail.com
Content-Type: text/plain; charset=UTF-8

We have Saturday lab coverage which includes IHC.
Case/slides vary between 8-15/25-85.  This includes multiplex and ISH.

Vikki
On Aug 23, 2015 10:49 AM, Cartun, Richard via Histonet  
histonet@lists.utsouthwestern.edu wrote:

 How many of you working in hospital-based pathology laboratories run 
 IHC on weekends?  Thank you.

 Richard

 Richard W. Cartun, MS, PhD
 Director, Histology  Immunopathology
 Director, Biospecimen Collection Programs Assistant Director, Anatomic 
 Pathology Hartford Hospital
 80 Seymour Street
 Hartford, CT  06102
 (860) 972-1596
 (860) 545-2204 Fax


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 the intended recipient(s) and may contain confidential and privileged 
 information. Any unauthorized review, use, disclosure, or distribution 
 is prohibited. If you are not the intended recipient, or an employee 
 or agent responsible for delivering the message to the intended 
 recipient, please contact the sender by reply e-mail and destroy all 
 copies of the original message, including any attachments.
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End of Histonet Digest, Vol 141, Issue 21
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Re: [Histonet] IHC Weekend Coverage

[Victoria Baker via Histonet]

We have Saturday lab coverage which includes IHC.
Case/slides vary between 8-15/25-85.  This includes multiplex and ISH.

Vikki
On Aug 23, 2015 10:49 AM, Cartun, Richard via Histonet 
histonet@lists.utsouthwestern.edu wrote:

 How many of you working in hospital-based pathology laboratories run IHC
 on weekends?  Thank you.

 Richard

 Richard W. Cartun, MS, PhD
 Director, Histology  Immunopathology
 Director, Biospecimen Collection Programs
 Assistant Director, Anatomic Pathology
 Hartford Hospital
 80 Seymour Street
 Hartford, CT  06102
 (860) 972-1596
 (860) 545-2204 Fax


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 intended recipient(s) and may contain confidential and privileged
 information. Any unauthorized review, use, disclosure, or distribution is
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 responsible for delivering the message to the intended recipient, please
 contact the sender by reply e-mail and destroy all copies of the original
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Re: [Histonet] IHC Weekend Coverage

[Kim Donadio via Histonet]

I never did on a routine basis. The only time was rare urgent cases or cases 
that needed done due to someone off or equipment problems. 

Kim D
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Re: [Histonet] IHC negative controls

[Shannon Gower via Histonet]

We purchased a Universal Negative reagent from Ventana and then we use that in 
a prep kit, totally eliminating the need for Mouse and Rabbit negatives and a 
lot of extra space on the carousel.

Shannon Gower
Senior Histotechnologist HTL(ASCP)
All Children’s Hospital
St. Petersburg, FL
(727-767-3058)
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Re: [Histonet] IHC negative controls

[Tim H via Histonet]

We run patient tissue as a negative and we use diluent as the reagent negative.
 
TH
 
 Message: 7
 Date: Fri, 17 Jul 2015 14:48:17 +
 From: Abbott, Tanya tanyaabb...@catholichealth.net
 To: histonet@lists.utsouthwestern.edu
   histonet@lists.utsouthwestern.edu
 Subject: [Histonet] IHC negative controls
 Message-ID:
   
 852f7d2c14fb464d80e182b15db138af6b7ec...@chiex005.chi.catholichealth.net
   
 Content-Type: text/plain; charset=us-ascii
 
 Happy Friday everyone! We currently have a Ventana Ultra. I am wondering what 
 everyone is doing/running for a negative reagent control and/or a negative 
 tissue control?
 Thanks in advance!
 Tanya
 
 Tanya G. Abbott
 Manager Technologist
 Histology/Cytology
 St Joseph Medical Center
 (phone) 610-378-2635
 
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Re: [Histonet] IHC repeat rate

[Terri Braud]

I once had a vendor tell me that 5% was an acceptable rate.  I am more inclined 
to use 2%

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874
   
4. IHC repeat rates (Bitting, Angela K.)
Message: 4
Date: Thu, 25 Jun 2015 15:08:46 +
From: Bitting, Angela K. akbitt...@geisinger.edu
Subject: [Histonet] IHC repeat rates

What does this group feel is an acceptable repeat rate for automated IHC 
staining??
Thanks for your responses in advance.
Angie
***


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Re: [Histonet] IHC repeat rate

[Blazek, Linda]

I would think that it would really depend on if it is a repeat due to an 
instrument problem or a human error.  I think that if a vendor says that a 5% 
repeat rate is acceptable for their instrument I would look for another vendor. 

-Original Message-
From: Terri Braud [mailto:tbr...@holyredeemer.com] 
Sent: Thursday, June 25, 2015 3:32 PM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC repeat rate

I once had a vendor tell me that 5% was an acceptable rate.  I am more inclined 
to use 2%

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874
   
4. IHC repeat rates (Bitting, Angela K.)
Message: 4
Date: Thu, 25 Jun 2015 15:08:46 +
From: Bitting, Angela K. akbitt...@geisinger.edu
Subject: [Histonet] IHC repeat rates

What does this group feel is an acceptable repeat rate for automated IHC 
staining??
Thanks for your responses in advance.
Angie
***


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Re: [Histonet] IHC repeat rates

[Tony Henwood (SCHN)]

The RCPA (in Australia) recently requested that laboratories perform Technical 
Failure Repeat Audits that might be of use.
Subsequently I produced a brief report for our Society's journal:

Audit of Technical Failures
Tony Henwood
Histopathology
The Children's Hospital at Westmead, Sydney
The RCPA has requested that laboratories implement the Internal Quality 
Assurance Framework-Pilot Program for the interpretative and morphological 
disciplines. There is also a requirement for 2.5 hours of 'other', internal 
quality activities relating to the technical and service measures section of 
the framework (over a 3 month timeframe). One of these is the re-staining 
required due to technical failure (HE, special stain, Immunoperoxidase, etc.). 
This Technical Failures Audit records the number of stains of a certain type 
requiring repeat over total number stained during a 3 month audit period. One 
would also include those instances where recuts or deeper sections are 
requested due to sectioning artefacts or incomplete full face.  The necessity 
for repeat staining reflects the quality of technical work and has impact on 
overall laboratory efficiency.
This audit is one of the measures recommended to monitor the analytic phase of 
the test cycle. The analytic phase of the test cycle represents all of the 
steps in processing and evaluating the specimen in the laboratory until a 
diagnosis is rendered (3). Other audits include (6):
*   Quality of histologic sections
*   Specimens lost in processing
*   Turnaround time (TAT)
*   Block labelling
*   Slide labelling
*   Extraneous tissue
*   TAT for immunohistochemical analysis
*   Report audit for integration of stains with morphologic diagnosis
*   Annual review of antibody inventory and frequency of use
The Association of Directors of Anatomic and Surgical Pathology have included 
Frequency and causes of repeated stains under Immunohistochemical analysis 
(6). But one would think that this could, and probably should, be applied 
across all the technical aspects of the analytical phase of histopathology.
An effective audit needs to be relevant, objective, quantified and repeatable 
with formal identification of areas requiring improvement (1, 2). It also 
provides a method for reassessment of performance once appropriate changes have 
been implemented (1).
Although errors causing patient harm are unfortunate, it must be acknowledged 
that a certain error rate is prevalent. Error levels that may be deemed within 
an acceptable range should be determined based on the literature for that 
measure, with the goal of modification by continuous improvement (6)
It has been recommended that a Quality Assurance Monitor should include a 
targeted acceptable range and reference benchmark data (3). It is appropriate 
to complete this audit, but it can be of limited use unless we have appropriate 
benchmarks to compare our laboratories data to. 
From the literature:
*   Zuk et al (1) found 19% of cases were technical unsatisfactory 
(holes, folds, debris, chatters, scores or absence of full transverse 
section). They also found that HE staining was poor in a 2% of cases. This 
study recorded all cases of technical failure, not just those requiring a 
repeat in order to facilitate a diagnosis.
*   In the manufacturing industry, six sigma has emerged as a generic 
quality standard. This standard aims to have the total number of failures in 
quality, or customer satisfaction at 3.4 defects or fewer than 4 defects per 
million products (4).
*   Morelli et al (5) found in their study of errors in histology 
preparation that 0.03% of cases required the repeat of one or more processes;
To implement this audit we also need to determine the Rules of Engagement- 
what parameters need to be observed in order to render the results meaningful?
*   Should a repeat caused by microscopic incomplete full face (not obvious 
from scrutinising the paraffin block) be counted?
*   Should further sections and in particular serials to improve the 
detection of a small focus of disease be counted as a technical failure?
So, what do you think?
Please send your thoughts to the editor
References:
1.  Zuk, J. A., Kenyon, W. E.,  Myskow, M. W. (1991). Audit in 
histopathology: description of an internal quality assessment scheme with 
analysis of preliminary results. Journal of clinical pathology, 44(1), 10-16.
2.  Shaw CD, Costain DW. 1989 Guidelines for medical audit: seven 
principles. Br Med J 299:498-9.
3.  Nakhleh, R. E. (2009). Core components of a comprehensive quality 
assurance program in anatomic pathology. Advances in anatomic pathology, 16(6), 
418-423.
4.  Nakhleh, R. E. (2006). What is quality in surgical pathology?. Journal 
of clinical pathology, 59(7), 669-672.
5.  Morelli, P., Porazzi, E., Ruspini, M., Restelli, U.,  Banfi, G. 
(2013). Analysis of errors in histology by root cause analysis: a pilot study. 
J 

Re: [Histonet] IHC turnaround time

[Sebree Linda A]

We have 2 FTEs in IHC; myself as one of two people that started the lab 20+ 
years ago and 2 histotechs that rotate through on a weekly basis.  Our monthly 
totals currently run between 2400 and 3300.  Our TAT policy is in by noon, out 
by 5pm except for dual and triple stains (need to be on an instrument by 11:00 
am) and EBER (needs to be on by ~ 9:00 am) and HER2 dual ISH (only run 
overnight).  We have 4 Roche (Ventana) Ultras that hold 30 slides each and are 
continuous access.  All instruments are used every day and at least one 
(usually 2 or 3) in use overnight.

Linda

Linda A. Sebree 
University of Wisconsin Hospital  Clinics 
IHC/ISH Laboratory, Rm A4/204-3224 
600 Highland Ave. 
Madison, WI 53792 
(608)265-6596 
FAX: (608)262-7174 


-Original Message-
From: Cartun, Richard [mailto:richard.car...@hhchealth.org] 
Sent: Friday, June 19, 2015 10:21 AM
To: Olszewski, Dawn; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC turnaround time

I direct a busy and very efficient (I think) IHC lab staffed with 2.875 FTEs 
who spend most of their time doing IHC, but they also help Histology embed and 
cut paraffin blocks when needed (which is most days).  For 2015 we are 
averaging 4,022 IHC patient test slides and 1,650 IHC control slides per month. 
 Our IHC lab is staffed from 4:30 AM to 6 PM (or later depending on workload).  
We have 5 Leica Bond Max platforms; however, we still do some IHCs on the 
bench.  We usually do 2-3 runs per day and an overnight run (except on Fridays) 
as well.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs Assistant Director, Anatomic 
Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax

-Original Message-
From: Olszewski, Dawn [mailto:dawn.olszew...@sgmc.org]
Sent: Friday, June 19, 2015 10:53 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC turnaround time

Hi Histonetters,
  6-19-15

We are wondering what the average turnaround times for IHC are for other labs.  
We use the Biocare Intellipath instrumentation. It holds up to 50 slides ( 10 
slides per 5 racks) with continuous feed per open rack.  All IHC orders placed 
by 12pm are usually out the same day by 5pm. We average 434 IHC slides per 
month and have a staff of 3 FTE's.

A pathologist has voiced concerns over our IHC output.  We are trying to 
determine best practices for IHC turnaround time as measured from time order 
placed to time of slide delivery.

If you could respond with your IHC TAT including number of techs and average of 
IHC slides monthly, we would appreciate any input you may be able to provide.

Thank you in advance,

Dawn Olszewski, HTL(ASCP)QIHC

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Re: [Histonet] IHC turnaround time

[Rene J Buesa]

TAT is variable but, as a rule, IF the case is slotted for IHC testing the 
slides will be delivered along with the HE early next day (if received after 
4PM) or by late afternoon the same day (if received before 10AM)René 


 On Friday, June 19, 2015 11:11 AM, Olszewski, Dawn 
dawn.olszew...@sgmc.org wrote:
   

 Hi Histonetters,                                                               
                                                                   6-19-15

We are wondering what the average turnaround times for IHC are for other labs.  
We use the Biocare Intellipath instrumentation. It holds up to 50 slides ( 10 
slides per 5 racks) with continuous feed per open rack.  All IHC orders placed 
by 12pm are usually out the same day by 5pm. We average 434 IHC slides per 
month and have a staff of 3 FTE's.

A pathologist has voiced concerns over our IHC output.  We are trying to 
determine best practices for IHC turnaround time as measured from time order 
placed to time of slide delivery.

If you could respond with your IHC TAT including number of techs and average of 
IHC slides monthly, we would appreciate any input you may be able to provide.

Thank you in advance,

Dawn Olszewski, HTL(ASCP)QIHC

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Re: [Histonet] IHC turnaround time

[Cartun, Richard]

I direct a busy and very efficient (I think) IHC lab staffed with 2.875 FTEs 
who spend most of their time doing IHC, but they also help Histology embed and 
cut paraffin blocks when needed (which is most days).  For 2015 we are 
averaging 4,022 IHC patient test slides and 1,650 IHC control slides per month. 
 Our IHC lab is staffed from 4:30 AM to 6 PM (or later depending on workload).  
We have 5 Leica Bond Max platforms; however, we still do some IHCs on the 
bench.  We usually do 2-3 runs per day and an overnight run (except on Fridays) 
as well.

Richard

Richard W. Cartun, MS, PhD
Director, Histology  Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax

-Original Message-
From: Olszewski, Dawn [mailto:dawn.olszew...@sgmc.org]
Sent: Friday, June 19, 2015 10:53 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC turnaround time

Hi Histonetters,
  6-19-15

We are wondering what the average turnaround times for IHC are for other labs.  
We use the Biocare Intellipath instrumentation. It holds up to 50 slides ( 10 
slides per 5 racks) with continuous feed per open rack.  All IHC orders placed 
by 12pm are usually out the same day by 5pm. We average 434 IHC slides per 
month and have a staff of 3 FTE's.

A pathologist has voiced concerns over our IHC output.  We are trying to 
determine best practices for IHC turnaround time as measured from time order 
placed to time of slide delivery.

If you could respond with your IHC TAT including number of techs and average of 
IHC slides monthly, we would appreciate any input you may be able to provide.

Thank you in advance,

Dawn Olszewski, HTL(ASCP)QIHC

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Re: [Histonet] IHC turnaround time

[STEVEN PINHEIRO]

Dawn,
TAT has many variables. We run mostly Leica Bond platforms, but do have some 
Ventana Benchmark access if needed. The average volume here about 125 slides a 
day. The available time is directly dependent or order protocols. If the order 
is available before 11 am, the slides will be available mid to late afternoon 
the same day. If the order is created between 11 am and 4 pm, the slides will 
be available late that same evening (but usually not reviewed by pathology 
until the next morning). Order placed after 4 pm are variable and may make the 
evening run but will most likely not be run until the next day. We do not have 
an overnight run.

Steve Pinheiro
708-327-2642


-Original Message-
From: Olszewski, Dawn [mailto:dawn.olszew...@sgmc.org] 
Sent: Friday, June 19, 2015 9:53 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC turnaround time

Hi Histonetters,
  6-19-15

We are wondering what the average turnaround times for IHC are for other labs.  
We use the Biocare Intellipath instrumentation. It holds up to 50 slides ( 10 
slides per 5 racks) with continuous feed per open rack.  All IHC orders placed 
by 12pm are usually out the same day by 5pm. We average 434 IHC slides per 
month and have a staff of 3 FTE's.

A pathologist has voiced concerns over our IHC output.  We are trying to 
determine best practices for IHC turnaround time as measured from time order 
placed to time of slide delivery.

If you could respond with your IHC TAT including number of techs and average of 
IHC slides monthly, we would appreciate any input you may be able to provide.

Thank you in advance,

Dawn Olszewski, HTL(ASCP)QIHC

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Re: [Histonet] IHC turnaround time

[Jay Lundgren]

 Let me get this straight, Dawn.  You are running In by 12, out by 5.
on IHCs and your pathologist is* complaining*?  A 5 hour TAT is awesome!
Most places I've worked (and I've seen a lot in 16 years as a traveling
tech) have at least a 24 hour TAT for IHC.

 I've got to get a shirt to the cleaners before 9 to get it back by 5!
Which is more complex, washing a shirt, or doing IHC on human tissue?!

  Sincerely,

   Jay A. Lundgren, M.S.,
HTL (ASCP)

On Fri, Jun 19, 2015 at 10:54 AM, STEVEN PINHEIRO spinhe...@lumc.edu
wrote:

 Dawn,
 TAT has many variables. We run mostly Leica Bond platforms, but do have
 some Ventana Benchmark access if needed. The average volume here about 125
 slides a day. The available time is directly dependent or order protocols.
 If the order is available before 11 am, the slides will be available mid to
 late afternoon the same day. If the order is created between 11 am and 4
 pm, the slides will be available late that same evening (but usually not
 reviewed by pathology until the next morning). Order placed after 4 pm are
 variable and may make the evening run but will most likely not be run until
 the next day. We do not have an overnight run.

 Steve Pinheiro
 708-327-2642


 -Original Message-
 From: Olszewski, Dawn [mailto:dawn.olszew...@sgmc.org]
 Sent: Friday, June 19, 2015 9:53 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] IHC turnaround time

 Hi Histonetters,

 6-19-15

 We are wondering what the average turnaround times for IHC are for other
 labs.  We use the Biocare Intellipath instrumentation. It holds up to 50
 slides ( 10 slides per 5 racks) with continuous feed per open rack.  All
 IHC orders placed by 12pm are usually out the same day by 5pm. We average
 434 IHC slides per month and have a staff of 3 FTE's.

 A pathologist has voiced concerns over our IHC output.  We are trying to
 determine best practices for IHC turnaround time as measured from time
 order placed to time of slide delivery.

 If you could respond with your IHC TAT including number of techs and
 average of IHC slides monthly, we would appreciate any input you may be
 able to provide.

 Thank you in advance,

 Dawn Olszewski, HTL(ASCP)QIHC

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 http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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 and is intended for the sole use of the intended recipient(s). It may
 contain information that is privileged and confidential.  Any unauthorized
 review, use, disclosure, or distribution is prohibited. If you are not the
 intended recipient, please delete this message, and reply to the sender
 regarding the error in a separate email.


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Re: [Histonet] IHC on fish

[Goins, Tresa]

Cate -

Your gut is probably right but results are unpredictable.
I have forwarded your query to the Bozeman Fish Technology center to see if 
they can offer any assistance.

Tresa

-Original Message-
From: Hardy, Cate [mailto:cha...@csu.edu.au] 
Sent: Sunday, June 14, 2015 4:10 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC on fish

Hi all;

Has anyone performed IHC on fish. I have been asked to perform this using 
several of our routine antibodies that I use on mammals. My gut tells me it 
won't work but I am a lowly histotech and don't know anything. Seeking 
knowledge from learned tech's with experience in such matters

Thanks

Cate Hardy
Senior Technical Officer
Veterinary Diagnostic Laboratory
Charles Strut University
NSW Australia
Charles Sturt University

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