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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[Histonet] IHC Validation Question

[Kara, Phillip via Histonet]

Morning Histo Friends.

Got a quick question for you. I am getting ready to go over to a new start up 
lab that will be doing all IHC staining in house.
I have set up a lab before and done validations but that was on equipment and 
special stains which I will be doing again and feel pretty good about it.
Any tips and tricks for validating IHCs? We will be using the Benchmark Ultra 
(please no solicitation we are all set).
We are hoping not to have to buy too many controls so I have been looking to 
try finding some in house I can use to help save the cost. If anyone has any 
good advice for this to I will be all ears.

Thanks in advance.

Phillip Kara
Senior Research Associate
UNT HSC
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[Histonet] IHC

[Chakib Boussahmain via Histonet]

Greetings Histonet community,

I'm interested in learning if anyone has experience withperforming Von 
Willebrand factor and alpha smooth muscle actin IHC staining onpig tissues 
(FFPE). If you do, could you please share the protocol along withdetails about 
the antibodies used, including ordering cat# and clones?

Any assistance you could provide would be immensely appreciated.

Best regards, 
Chakib
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[Histonet] IHC/Special staining

[Ken M via Histonet]

Does anyone know of any lab with excess capacity good quality and fast 
turnaround that could do IHC/special staining on the side? We are out of 
capacity, and we don't always have the antibodies that we need.

Kevin
kdea...@hotmail.com
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[Histonet] IHC CRO

[Amy Lee via Histonet]

Hello,
Does anyone has a list of category can share that is used for IHC CRO
comparison? I want to make a table to do comparison after talking to
multiple CROs and I don't want to miss something.  Just wondering if anyone
did this before and can share your table or list?

Thanks,
Wendy
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[Histonet] IHC wash buffer temperature?

[Preiszner, Johanna via Histonet]

Hello Histonetters,

I work in research and we get too much non-specific background on our IHC-s. 
Anybody tried using wash solution at 37C instead of at room temperature?
If not, why? Is it a bad idea?
Thanks,
Hanna Preiszner
ETSU Pathology
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[Histonet] IHC QC Forms

[ADESUPO ADESUYI via Histonet]

Hi,

I will be glad if you guys can share your IHC QC forms with me.

Sincerely,
Adesupo A


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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
> for transmission to (or receipt by) any unauthorized person(s). If you
> believe that you have received this email in error, do not read any
> attachments. Instead, kindly reply to the sender stating that you have
> received the message in error. Then destroy it and any attachments. Thank
> 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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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 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 
display address in the From: field. Avoid action unless you know the content is 
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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[Histonet] IHC gunk

[Karen Heckford CA-San Francisco via Histonet]

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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notify us  immediately by reply email.  Thank you

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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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[Histonet] IHC staining of tendons and cartilage

[Charles Riley via Histonet]

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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[Histonet] IHC systems

[warda hassan via Histonet]

Dear all




We are planning to purchase a new IHC system.

May I request your valuable feedback, observation and troubleshooting
encountered while handling Leica Bond 3?

Would you prefer Leica or Roche for the quality of staining, consumables
supply and user-friendly lean management with help in TAT improvement?

Kind Regards


Thanking you for helping out
W Anwar
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[Histonet] IHC position in Gainesville FL

[Dooley, Elaine O. via Histonet]

Hi Histonetters,

Just wanted to let everyone know I am looking for an histotechnologist to work 
in the IHC area of the Histology lab here at UF Health Shands hospital.
I am looking for an a person that is energetic, hardworking and willing to 
learn all aspects of the immunohistochemistry area.

The Gainesville area has a lot to offer being a college town.  Also there is 
world class springs for scuba diving, and rivers for kayacking and many nature 
trails.
The coast is only a couple of hour to either side if the beach is your thing.

Please check out our posting 
https://shands.careers.ufhealth.org/jobs/Histologist_Histo/Surgical_/Gainesville_Florida/100/314081/


If you want to talk about the position with me personally call 352-265-0111 ext 
72117
Elaine Dooley IHC lead tech
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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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and destroy any printouts.

-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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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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[Histonet] IHC background

[Paula via Histonet]

Hello everyone,

 

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

 

Thank you in advance.

Paula

Bio-Path Medical Group

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[Histonet] IHC staining of cartilage

[Bacon, Charles via Histonet]

We use P8920 Sigma-Aldrich Poly-L-lysine solution. Great retention without 
excess background staining. You can prep a rack of slides in diluted solution 
but we put a gloved finger over the end of the bottle and and swipe it down the 
slide at full strength. Let the slide dry and move forward as you normally 
would. 

Chuck Bacon, HTL(ASCP)CM 
Supervisor Histology
Baystate Medical Center
361 Whitney Ave., Holyoke, MA 01040
Telephone: 413-322-4786  Fax: 413-322-4790
charles.ba...@baystatehealth.org

-Original Message-
From: histonet-requ...@lists.utsouthwestern.edu 
 
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To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 220, Issue 2

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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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[Histonet] IHC staining of cartilage

[Charles Riley via Histonet]

I recently tried running an automated ihc test on a few pieces of cartilage
tissue.

All of the sections fell off my slides. I am using the TOMO charged slides
and have never had any issues.

Do cartilage sections require longer drying times in order for the tissue
to adhere better?
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[Histonet] IHC Research Technician Kansas State Veterinary Diagnostic Lab

[Jennifer Phinney via Histonet]

Hello HistoNetters,
The Kansas State Veterinary Diagnostic Laboratory (KSVDL) within the College of 
Veterinary Medicine is seeking applicants for a Research Assistant. This 
position will have the primary responsibility of coordinating new incoming 
research projects and developing new tests related to immunohistochemistry. A 
large portion of this person's time will be spent developing new diagnostic and 
research assays along with coordinating research projects. This individual will 
also spend a decent amount of time performing diagnostic immunohistochemistry 
assays and will also assist in diagnostic routine histology duties related to 
service and research.

KSVDL is an AAVLD accredited lab.

If anyone is interested in applying, the link to the job posting may be found 
here:
https://careers.k-state.edu/cw/en-us/job/510780/research-assistant-histology-veterinary-diagnostic-laboratory

Jennifer Phinney MS, QIHC
Histology Laboratory Administrator
KSVDL
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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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[Histonet] IHC with zamboni's fixed tissues

[Colleen Forster via Histonet]

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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[Histonet] IHC Disclaimer for immunofluorescence stains

[Morken, Timothy via Histonet]

Those labs doing diagnostic immunofluorescence for kidney or skin, do you put 
in the IHC disclaimer used for peroxidase staining on the reports?

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

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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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[Histonet] IHC validation after a new tissue processer is installed

[Martha Ward-Pathology via Histonet]

Hello all,

After a very long time our histology department is getting a new tissue 
processor, and hopefully several more in the near future.   Of course for IHC 
that means a revalidation/reverification of our IHC stains.  Since I have not 
had to do this before I wanted to get some guidance on how to go about doing 
this.  I feel like I have to completely revalidate the ER, PR and Her2 (20 
positive, 20 negative) but wasn't sure about the other antibodies.   We have a 
menu of around 130do I have to test every one of them or can we chose a 
representative sample? How many antibodies would you suggest?   How many 
positive and negative cases of each should we run?   It says it is left up to 
the medical director but with our CAP inspection next spring we want to make 
sure we will be fully prepared.What have others done?

Thank you in advance for your help with this.


Martha Ward, MT (ASCP) QIHC
Wake Forest Baptist Medical Center
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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

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.




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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[Histonet] IHC validations

[Paula via Histonet]

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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[Histonet] IHC validation

[Kristy Castillo via Histonet]

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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[Histonet] IHC control blocks, DAB kit

[Cassie P. Davis via Histonet]

Kristy, this might be of some help too: 
https://www.fixationonhistology.com/post/control-tissue-faqs
[https://static.wixstatic.com/media/128854_2fda1d8f8d5c48cdad7701b3e866d885~mv2.jpg/v1/fit/w_1000,h_728,al_c,q_80/file.png]
Control Tissue FAQs - 
fixationonhistology.com
Should control slides be refrigerated? Some controls such as placenta, p53, and 
p57, for example, may lose antigenicity (the antigen’s ability to bind with the 
products of the cell mediated response, like B-cells and T-cells), if they are 
left out. Others are fine being left on the shelf. Whether or not you need to 
refrigerate them also depends on how far in advance they have been cut.
www.fixationonhistology.com


Cassie Davis
Histology Department
Anatomical Pathology Laboratory
302-575-8095
Email:  cda...@che-east.org

W  302-575-8095

701 North Clayton St.
Wilmington, DE 19805

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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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> Histonet@lists.utsouthwestern.edu
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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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[Histonet] IHC DAB

[Kristy Castillo via Histonet]

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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[Histonet] IHC question

[Blanca Lopez via Histonet]

I have a question regarding ihc
I’m running some ihc using Dako linker 48, this gave me a notice of short 
reagent but when I made it my time was off and suspended. The slides are just 
being on peroxide block for 5 min.
Can I just reload them again? Is this will not affect my staining
Ir what is you suggestion
Thanks

Blanca Lopez HT(ASCP)
Histotechnologist
UT Southwestern Medical Center
Harold C Simmons Comprehensive Cancer Center
Pathology Lab NB5-102
214-648-7598
blanca.lo...@utsouthwestern.edu




UT Southwestern


Medical Center



The future of medicine, today.

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[Histonet] IHC Troubleshooting and Water Baths

[Nancy Schmitt via Histonet]

Hi Charles-

IHC staining - We also use the Leica Bond staining system and had tissue fall 
off in the beginning.  We blot the slides to remove any additional water and 
then place in drying oven for 30 minutes at 60 before placing on the stainer.

Water baths - at the end of each day the water is dumped, the bath is rinsed 
with bleach solution, rinsed with water and dried.

Nancy Schmitt MLT, HT(ASCP)
Pathology Support Services Manager


-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


Message: 5
Date: Wed, 26 Feb 2020 17:09:14 -0500
From: Charles Riley 
To: Histo List 
Subject: [Histonet] Poll question
Message-ID:

Content-Type: text/plain; charset="UTF-8"

How often does everyone clean their water-bath?

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs



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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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[Histonet] IHC troubleshooting

[Charles Riley via Histonet]

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<mailto:l...@premierlab.com>
www.premierlab.com<http://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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[Histonet] IHC on Pig Tissues

[Lori Sorrells via Histonet]

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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[Histonet] IHC Application Specialist Job in Boston-

[Melissa Owens via Histonet]

Hello and Happy Monday,

I have an "off the bench" position available for an IHC Application Specialist. 
Looking for someone with extensive IHC experience and a research emphasis 
rather than clinical emphasis on experience. Bachelor's Degree at least. 
Ability to travel throughout Boston and the Northeastern States. 75% travel 
involved. Must be in Boston area or open to relocate to the Boston area due to 
the primary customer base in Boston. Message me directly for more details!

Melissa Owens
Allied Search Partners
AN MRINETWORK MEMBER
Direct (Call) Line:  386.265.1368
Text Me: 386.855.8758
Toll Free: 888.388.7571 ext. 102
Fax: 888.388.7572

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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<mailto: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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> Histonet@lists.utsouthwestern.edu
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>
> ___
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> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


-- 
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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[Histonet] IHC mouse optomization

[Blanca Lopez via Histonet]

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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[Histonet] IHC Specialists Needed CA, IL, AZ and New England.

[Pam Barker via Histonet]

Hi Histonetters!
Just a quick note about some exciting new opportunities.
I am recruiting for IHC Specialists nationwide.
I have a number of roles ranging from in-house to field applications
specialists.
If you are interested please shoot me a quick email for details.
And. If you refer someone that I place you will earn a referral fee.

Thanks-Pam

To subscribe to my histology careers bulletin reply to this email with
subscribe.
#jobs4myhistopeeps 
#ilovemyhistopeeps
Follow my hashtags and make your day great and your career greater!!

Right Place, Right Time, Right Move with RELIA!

Thank You!
 Pam M. Barker 
Pam Barker
President/Senior Recruiting Specialist-Histology
RELIA Solutions
Specialists in Allied Healthcare Recruiting
5703 Red Bug Lake Road #330
Winter Springs, FL 32708-4969
Phone: (407)657-2027
Cell: (407)353-5070
FAX: (407)678-2788
E-mail: rel...@earthlink.net 
https://www.facebook.com/RELIASolutionsforhistologyprofessionals
www.facebook.com  /PamBarkerRELIA
www.linkedin.com/in/reliasolutions
www.twitter.com/pamatrelia 





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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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think you have received this message in error, please advise the sender and
then delete this message and any attachments immediately.*




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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[Histonet] IHC Control Slide Longevity

[Sandra Cheasty via Histonet]

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<mailto: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<mailto:richard.car...@hhchealth.org>


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[Histonet] IHC Billing - 88342 vs. 88341

[Cartun, Richard via Histonet]

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


This e-mail message, including any attachments, is for the sole use of 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 
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Re: [Histonet] Histonet - IHC Manual staining

[Colleen Forster via Histonet]

Hello Dave,

I still do a lot of manual work and I also use Biocare reagents. They work
well and their customer service is very good.

Colleen Forster

On Fri, Jan 18, 2019 at 12:58 PM Jeanine Ronkowski via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> Hi Dave:Our lab has used reagents from Biocare Medical for a very long
> time. They’re packaged in 6ml and 25 ml vials and ideal for hand staining.
> And their technical support is great tooJeanine
>
> Hello Histoneters -
>
> Does anyone still do manual IHC's? If so, which IHC staining kits would
> you recommend? Thanks.
>
> Yours,
> Dave
>
> Sent from Yahoo Mail for iPhone
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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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[Histonet] Histonet - IHC Manual staining

[Jeanine Ronkowski via Histonet]

Hi Dave:Our lab has used reagents from Biocare Medical for a very long time. 
They’re packaged in 6ml and 25 ml vials and ideal for hand staining. And their 
technical support is great tooJeanine

Hello Histoneters -

Does anyone still do manual IHC's? If so, which IHC staining kits would you 
recommend? Thanks.

Yours,
Dave

Sent from Yahoo Mail for iPhone
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[Histonet] IHC Manual Staining

[David via Histonet]

Hello Histoneters -

Does anyone still do manual IHC's? If so, which IHC staining kits would you 
recommend? Thanks.

Yours,
Dave


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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


*Everything in moderation...even moderation itself**!*
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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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[Histonet] IHC-H HELP...

[Curt via Histonet]

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
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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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<http://aka.ms/weboutlook> 
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[Histonet] IHC Negative reagent controls

[Cayman Fleck via Histonet]

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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[Histonet] IHC control slides for EBNA

[Michele Wich via Histonet]

Does anyone know of a source of IHC control slides for EBV Nuclear Antigen?

Thank you.
Michele



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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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[Histonet] IHC Stainer

[Sandra Cheasty via Histonet]

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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[Histonet] IHC System from StatLab

[Jeanine Ronkowski via Histonet]

I’m pleased to report that I was finally contacted by a representative from 
StatLab that provided me with the information I was seeking.  I realize that 
their system may not be fully released, but I didn’t think it would be that 
difficult for them to provide information.  Turns out that this instrument is 
very similar to another manufacturer’s instrument that’s been on the market for 
a while.J


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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

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[Histonet] IHC System from StatLab

[Jeanine Ronkowski via Histonet]

I’ve heard from a number of people that Stat Lab plans to release a new 
instrument for immunostaining and I was wondering if someone, including the 
folks at StatLab, might be able to provide more information.Thx,Jeanine


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[Histonet] IHC question

[Charles Riley via Histonet]

Does anyone run this antibody on their Leica bond platforms?


BRAF V600E (VE1)?  I am looing to purchase it from sigma aldrich
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[Histonet] IHC Symposium in Quebec City

[John Garratt via Histonet]

2018 cIQc Symposium: IHC Toolkits for Diagnostic Pathology

Where:  Quebec City at the  Hotel Palace Royale (775 Boulevard
Honoré-Mercier)

When: September 20-21, 2018.

Go to   www.ciqc.ca  for more information.

 

The symposium is for Lab Scientists, Pathologists and Industry to discuss a
variety of topics including PD-L1, Alk, Ros1 

 

John Garratt – Canadian Immunohistochemistry Quality Control

 

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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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[Histonet] IHC and Histogel

[Paula via Histonet]

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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[Histonet] IHC Platforms

[Campbell, Tasha M. via Histonet]

Hello everyone.  I was wondering if any small, GI labs do IHC's in house and if 
so what platforms you use.  I am just researching right now to see if it would 
be worth it to do in house or not.  I know a lot of these instruments and 
reagents are expensive so I would be looking for the most inexpensive 
instrument and reagents without sacrificing quality.  Thanks in advance for 
your help!!!






Tasha Campbell, B.S.,HTL(ASCP)
Frederick Gastroenterology Associates
310 W. 9th St.
Frederick, MD 21701
301-695-6800 ext. 144

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[Histonet] IHC protocol help

[Charles Riley via Histonet]

Is anyone out there using the Leica Bond III and Bond Max to perform the
following tests?

ACD RNA ISH Probe EBV from Leica


CDK4 and or MDM2 from ABCAM

Amyloid P from Biocare

Amyloid A from Epitomics

I am trying to optimize and validate them our on machines  and would like
some help where to start. If you don't mind sharing your protocol it would
be greatly appreciated. Or if you know of another source for any of these
antibodies you think is better please let me know.

Thank you in advance

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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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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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



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