[Histonet] Response to Considering whether starting your own repository due to high quantity of tissues already in your posession that are being discarded and alternately used for profitable gain

[Eddie Martin via Histonet]

Hello Curt,

Regarding your question:
So running a ref lab for years I've acquired a rather large supply of old
blocks of various tissues, I'm considering turning this into a tissue
bank/repository with the purpose of selling blocks for research and control
purposes... anyone have any experience, what are the requirements to be
compliant??? I have zero experience with this. I'm guessing some release
forms, maybe patient consent forms (IMPOSSIBLE TO GET THOSE FOR BLOCKS THAT
ARE 10 YRS OLD)... release of liability forms... I'm starting at zero so
any insight is appreciated!


*Note 1: I'm neither a pathologist, a lawyer or any kind of adequate legal
counsel. The response below is my opinion only, and my own understanding of
the HIPAA act, and does not reflect the views of the institutions I work at
or the employers I work for:

I kindly suggest you seek permission from your company's medical-legal
office, as they and not yourself individually are covered entities. Any
instance of accessing a patient's medical diagnostic report for non-testing
purposes that is also not assigned to you by a clinician/pathologist of
accessing medically necessary information to perform tasks covered and
protected by the covered entity, including testing necessary to perform for
the patient's medical care and continuation of care is a breach of a
patient's PHI information, whether accessed via hardcopy or electronic or
any other kind of format.  I can easily think of four Federal Acts in place
protecting PHI:

** HIPAA, HITECH, FERPA, GINA

*** Then individual State Laws that each may have even more strict
guidelines in place to protect PHI.

I get that you are trying to repurpose pathology materials, but established
repositories are regulated by the FDA in addition to other federal and
State agencies. Seeking the permission of your company's medical-legal
department is highly suggested before accessing anything. My understanding
of the intricacies of what I'm posting is limited in scope and
understanding. From my understanding of the HIPAA Act above, each instance
of breach must be reported to your company within sixty days of becoming
aware of the breach, unless your local and State authorities have more
stringent laws in place; and may subject your employer and yourself to
breaching the confidentiality, integrity and availability made available in
accessing the PHI, that also outlines standards and penalties for
non-compliance.

In the event the quantity of breaches exceeds 500 individuals impacted, the
covered entity must also report the breach to the Department of Health and
Human Services and provide the information to prominent media outlets to
inform possible patients impacted by the breach within the jurisdiction the
breach had occurred.

Very respectfully,

Eddie Martin, HT, HTL, QIHC


On Thu, Aug 1, 2024 at 1:00 PM 
wrote:

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> Today's Topics:
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>1. Tissue repository (Curt Tague)
>2. Re: Tissue repository (Jay Lundgren)
>3. Re: Tissue repository (richard cartun)
>4. Re: Tissue repository (Curt Tague)
>5. Happy Thursday Histonetters!! (rel...@earthlink.net)
>
>
>
> -- Forwarded message --
> From: Curt Tague 
> To: "Histonet@lists.utsouthwestern.edu"  >
> Cc:
> Bcc:
> Date: Wed, 31 Jul 2024 21:02:20 +
> Subject: [Histonet] Tissue repository
> Hi all,
>
> So running a ref lab for years I've acquired a rather large supply of old
> blocks of various tissues, I'm considering turning this into a tissue
> bank/repository with the purpose of selling blocks for research and control
> purposes... anyone have any experience, what are the requirements to be
> compliant??? I have zero experience with this. I'm guessing some release
> forms, maybe patient consent forms (IMPOSSIBLE TO GET THOSE FOR BLOCKS THAT
> ARE 10 YRS OLD)... release of liability forms... I'm starting at zero so
> any insight is appreciated!
>
> Best!
>
> Curt
>
>
>
>
>
> -- Forwarded message --
> From: Jay Lundgren 
> To: Curt Tague 
> Cc: "Histonet@lists.utsouthwestern.edu"  >
> Bcc:
> Date: Wed, 31 Jul 2024 17:26:44 -0500
> Subject: Re: [Histonet] Tissue repository
> Sorry Igor, but if you can't get informed consent, it's not legal or
> ethical to sell patients' surgical specimens.  There are laws in all 50
> states. The body or parts thereof have to be explicitly donated.
>
> If you have informed consent, you can sell them for a large profit, as lots
> of non-transp

Re: [Histonet] IF staining questions

[Eddie Martin via Histonet]

I'd like to preface that I haven't done any Alexaflour testing in just over 10 
years. 
The simplest response may prefer using a conjugated secondary if you're 
planning to do multiplex or multicolor analysis. 

If I was limited in commercially available options for primaries needed for the 
test sample, then i would prefer a conjugated secondary as it loosens up 
testing requirements for testing. 
Another benefit is that it may increase specific testing and allow positive 
staining to appear more visibly present than a directly conjugated antibody. 

There are other benefits for a conjugated secondary or tertiary antibody, but 
it requires users to know how to troubleshoot the stain. 
Direct conjugation is easier to use in many circumstances. I would communicate 
to your friend to try both for the study and determine as a team of internal 
collaborators on which method is easiest and adequate enough for performing the 
method testing. 

I hope this helps. 
Eddie

NIH Bone Marrow Service 
RND clinical & anatomic pathology specialist

> On Oct 19, 2023, at 9:22 AM, Charles Riley  wrote:
> 
> Why would one decide to use a primary antibody along with a secondary
> antibody rather than a primary antibody conjugated with the secondary?
> 
> Example.   I have a researcher who wants to do CD11C staining with
> Alexafluor488
> 
> Is it better to buy and use a primary antibody CD11C conjugated with
> Alexafluor488  or to do the CD11C primary and a Rabbit anti-rat (H_L) IgG
> antibody secondary?
> 

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Re: [Histonet] Histonet Digest, Vol 237, Issue 4

[Eddie Martin via Histonet]

Thought on alternative for Sudan Black.  I don't use this stain...an
alternative is an Oil Red O stain.  Oil Red O is done on frozen
sections...but you can also deparaffinize FFPE sections to water, and then
perform your Oil Red O stain.

I hope this helps.

Very Respectfully,
Eddie Martin

Eddie Martin, HTL, QIHC
The National Institutes of Health
Bone Marrow Service
10 Center Drive
Building 10, Room 2C360
Bethesda, MD 20892
Office: 301-594-2054


On Thu, Aug 10, 2023 at 12:59 PM 
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>2. Sudan Black B (Betsy Molinari)
>
>
>
> -- Forwarded message --
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> To: 
> Cc:
> Bcc:
> Date: Wed, 9 Aug 2023 14:13:39 -0400
> Subject: [Histonet] Happy Hump Day Histopeeps!! Summer is Almost OVER!!!
> Here are some exciting new opportunities for you and your friends!
> Happy Hump Day Histopeeps!!
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> -- Forwarded message --
> From: Betsy Molinari 
> To: Histonet 
> Cc:
> Bcc:
> Date: Thu, 10 Aug 2023 14:57:31 +
> Subject: [Histonet] Sudan Black B
> Hi,
> I have been asked to do a Sudan stain on a heart  biopsy for lipofuscin.
> The biopsy is in a paraffin block. They are looking  to better report and
> understand the IHC. I am totally unfamiliar with this stain. I did some
> reading but have been unable to find a protocol for paraffin sections. I
> found a reference to Sheehan & Hrapchak (1973) but unfortunately I don't
> have that edition.  Any ideas would be greatly appreciate

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
>
>
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> -- 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&White 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
>
>
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>
> Good Morning,
> I have recently developed a problem with contamination of some kind on my
> IHC slides.
> The contamination is black clumps and lays on top of the tissue.  I have
> been told it is bacteria but the Pathologist and I kind of doubt that.   I
> cannot 

Re: [Histonet] Histonet Digest, Vol 232, Issue 7

[Eddie Martin via Histonet]

Hi kdea...@hotmail.com,

What likely is occurring but you didn’t mention in the thread is that your
friend is using alkaline phosphatase based detection, which, by itself can
react with kidney tissue. This likely explains why your friend is getting
strong kidney staining and weak melan-a staining.

This also tells me your friend didn’t optimize the antibody correctly. I’m
not sure why your friend is choosing to use a lyophilized antibody for
Melan-A as so many other commercially available options are available
capable of refrigerated storage and good up to 3 years. CellMarque makes an
Melan-A in both realty to use and/or a liquid concentrate with a 3-year
expiry date.

Best wishes,
Eddie Martin, HTL,QIHC
The National Institutes of Health
Department of Laboratory Medicine
Bone Marrow Service
eddie.mar...@nih.gov
(301)-594-2054



On Wed, Mar 15, 2023 at 1:00 PM 
wrote:

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>1. Problems with Melan A staining (Ken M)
>
>
>
> -- Forwarded message --
> From: Ken M 
> To: "histonet@lists.utsouthwestern.edu"  >
> Cc:
> Bcc:
> Date: Wed, 15 Mar 2023 16:51:03 +
> Subject: [Histonet] Problems with Melan A staining
> One of my histotech friends is having issues with Melan A staining. Using
> MA5-27949 Melan A antibody from Thermofisher, the negative Kidney tissue
> control was stained strongly positive while the positive melanoma tissue
> control was stained weakly positive. He couldn’t figure out why after
> several tries to reduce background. Can someone give me some suggestions?
> Thank you very much!
>
>
> Ken
>
>
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[Histonet] Suggest a Temperature Monitoring System

[Eddie Martin via Histonet]

Hi Matthew, 

Our laboratory performs clinical testing that supports the NIH Clinic Center. 
The current temperature monitoring system we’re using is acquired through 
ReesScientific.com 

The software is easy to use regardless of the extent of privileges a user has. 
The easiest advantage over freshloc I can think of is an option to find tagged 
equipment on a map within the lab. That feature is more helpful on 2nd and 3rd 
shift, or in the event a lab section is being renovated and equipment isn’t in 
its usual location. 

Best regards.
Very respectfully,
Eddie Martin

> On Jan 18, 2023, at 11:41 AM, Matthew D. Roark  wrote:
> 
> Good morning all!
> 
> Is there a wireless temperature monitoring system that you would recommend?
> 
> 
> 
> 
> 
> Matthew Roark, HTL(ASCP)CM
> Histology Specialist
> Laboratory
> 
> P 573-331-3982 | M 573-979-1925 | F 573-331-5049
> mro...@sfmc.net
> 
> Saint Francis Healthcare System
> 211 Saint Francis Drive
> Cape Girardeau, MO 63703
> 
> [https://www.sfmc.net/images/SFHC_logo_email_signature.png]
> 
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Re: [Histonet] Histonet Digest, Vol 225, Issue 14

[Eddie Martin via Histonet]

Changing your IHC slides to another brand with stronger slide adhesion is
suggested when working with bone and very fatty tissue to remain on the
slide.  shaking off as much moisture from the slide rack before placing
them to dry in the oven is another.  Some animal tissues benefit from
leaving in the oven overnight at 60 degrees celsius.

regarding slides with better tissue section adhesion, our bone marrow
pathology laboratory likes using the Azer Scientific's EMS200W+ charged
slides. There's another brand that's great for human brain, breast and also
animal tissues: I believe they're called Autofrost by Cancer diagnostics
(but I'm not sure if it's these, as the slides were used mostly at another
laboratory I used to work at five years ago).  I hope this all helps!

Best regards,
Eddie Martin, HTL, QIHC
Bethesda, MD

On Wed, Aug 31, 2022 at 1:00 PM 
wrote:

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> Today's Topics:
>
>1. Automated ihc staining of bone (Charles Riley)
>2. Re: Automated ihc staining of bone (Cooper, Brian)
>3. Information on the resistance/viability of different
>   tissues/cells to different freezing temperatures? (Alida Bailleul)
>
>
>
> -- Forwarded message --
> From: Charles Riley 
> To: histonet@lists.utsouthwestern.edu
> Cc:
> Bcc:
> Date: Tue, 30 Aug 2022 15:50:18 -0400
> Subject: [Histonet] Automated ihc staining of bone
> I am working on trying to get IHC stains optimized on bone samples of rat
> tibias. The issue I am running into is that the periosteum is
> separating from the bone marrow and wrinkling up over top of the rest of
> the sample.
>
> If anyone has any techniques to prevent this wrinkling/detachment of tissue
> from the main section during automated ihc staining it would be greatly
> appreciated.
>
>
>
>
> -- Forwarded message --
> From: "Cooper, Brian" 
> To: Charles Riley , "histonet@lists.utsouthwestern.edu" <
> histonet@lists.utsouthwestern.edu>
> Cc:
> Bcc:
> Date: Tue, 30 Aug 2022 20:09:14 +
> Subject: Re: [Histonet] Automated ihc staining of bone
> Charles,
>
> Try and blot your sections dry before placing them into the oven.  After
> picking up your section, place your slide onto a flat surface and take a
> paper towel or piece of filter paper, and gently press it down on top of
> the section to wick away all of the moisture.  We use this technique
> frequently here on bone marrow cores.
>
> Thanks,
>
> Brian D. Cooper, HT (ASCP)CM QIHC| Histology Supervisor
> Department of Pathology and Laboratory Medicine
> Children's Hospital Los Angeles
> 4650 Sunset Blvd MS#43- Los Angeles, CA 90027
> bcoo...@chla.usc.edu
>
> -Original Message-
> From: Charles Riley via Histonet 
> Sent: Tuesday, August 30, 2022 12:50 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Automated ihc staining of bone (EXTERNAL EMAIL)
>
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> I am working on trying to get IHC stains optimized on bone samples of rat
> tibias. The issue I am running into is that the periosteum is separating
> from the bone marrow and wrinkling up over top of the rest of the sample.
>
> If anyone has any techniques to prevent this wrinkling/detachment of
> tissue from the main section during automated ihc staining it would be
> greatly appreciated.
> ___
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Re: [Histonet] Leica Bond Red kit

[Eddie Martin via Histonet]

Hello all,
Yes, the Leica Bond and Bond Max has an Alk Phos detection kit available.
Though the red can bleed if left in alcohol, it is more stable than other
branded red detection kits. The Red kit is also very stable for long term
storage and exceedingly better than the competitor brands that also provide
an automated IHC solution.

In lieu of dehydrating, consider drying them in an oven post rinse to avoid
the dehydration steps prior to clearing and mounting.

Cheers,
Eddie Martin
The National Institutes of Health
Bone marrow pathology section
Bethesda, MD 20892

On Fri, Jun 10, 2022 at 1:00 PM 
wrote:

> Send Histonet mailing list submissions to
> histonet@lists.utsouthwestern.edu
>
> To subscribe or unsubscribe via the World Wide Web, visit
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
> histonet-requ...@lists.utsouthwestern.edu
>
> You can reach the person managing the list at
> histonet-ow...@lists.utsouthwestern.edu
>
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
> Today's Topics:
>
>1. Red chroamgen/LeicaBond andBondMax (John O?Brien)
>2. Re: Red chroamgen/LeicaBond andBondMax (Tim Morken)
>3. Re: Red chroamgen/LeicaBond andBondMax (Cartun, Richard)
>
>
>
> -- Forwarded message --
> From: "John O’Brien" 
> To: histonet@lists.utsouthwestern.edu
> Cc:
> Bcc:
> Date: Thu, 9 Jun 2022 10:21:04 -0700
> Subject: [Histonet] Red chroamgen/LeicaBond andBondMax
> Can some tell me if the Leica IHC Bond or Bond Max can preform the Red
> Chroamgen IHC stain?
> John
> IMEB
>
>
>
>
>
> -- Forwarded message --
> From: Tim Morken 
> To: "John O’Brien" 
> Cc: histonet@lists.utsouthwestern.edu
> Bcc:
> Date: Thu, 9 Jun 2022 10:58:47 -0700
> Subject: Re: [Histonet] Red chroamgen/LeicaBond andBondMax
> John,  yes, it is their Bond Refine Red detection kit
>
> Tim morken
>
> Sent from my iPhone
>
> > On Jun 9, 2022, at 10:23 AM, John O’Brien via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
> >
> > Can some tell me if the Leica IHC Bond or Bond Max can preform the Red
> Chroamgen IHC stain?
> > John
> > IMEB
> >
> > ___
> > Histonet mailing list
> > Histonet@lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
>
> -- Forwarded message --
> From: "Cartun, Richard" 
> To: "John O’Brien" 
> Cc: "histonet@lists.utsouthwestern.edu"  >
> Bcc:
> Date: Thu, 9 Jun 2022 18:18:55 +
> Subject: Re: [Histonet] Red chroamgen/LeicaBond andBondMax
> Yes; we run IHC tests everyday on our Bonds with Leica's "Red" detection
> kit.  You do need to be careful when dehydrating the slides prior to
> coverslipping since the chromogenic product is not as stable as DAB.
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & The Martin M. Berman, MD
>   Immunopathology/Morphologic Proteomics Laboratory
> Assistant Director, Anatomic Pathology
> Department of Pathology & Laboratory Medicine
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 972-1596 Office
> (860) 545-2204 (Fax)
>
>
>
> -Original Message-
> From: John O’Brien via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Thursday, June 9, 2022 1:21 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Red chroamgen/LeicaBond andBondMax
>
> CAUTION: This email is from outside HHC. USE CARE when opening attachments
> or links.
>
> Can some tell me if the Leica IHC Bond or Bond Max can preform the Red
> Chroamgen IHC stain?
> John
> IMEB
>
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>
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Re: [Histonet] Vol 203, Issue 21

[Eddie Martin via Histonet]

Hi Warda,

1. Both Roche and Leica have different instruments that are both equally
capable of breaking down.  It's really not feasible to determine which will
cost more in repairs on this premise alone.
2. Roche Ultra stains their probes with either a blue chromogen or you may
also use a Silver chromogen.

I hope this helps!
Eddie Martin

On Fri, Oct 30, 2020 at 3:30 AM warda hassan  wrote:

> Dear Eddie
>
> Thank you so much for your detailed explanation.
> Its really helpful, I just had two questions since you had worked on both
> the systems
>
> 1- Which one of them requires more maintenance and causes more breakdowns.
>
> 2- Does Roche performs FISH or is it SISH with probe IHC principle
>
> Thank you so much
> W Hassan
>
> On Fri, 30 Oct 2020 at 1:12 AM Eddie Martin  wrote:
>
>> Hi Histonet and W.A. Hassan,
>>
>> Supposing you have a slide volume of 15-20k slides per year & currently
>> use a Dako instrument, but would like feedback on whether to get a:
>>   * Bond Max, or
>>   * Roche Ultra
>>
>> My response would depend more than what your yearly slide volume is.
>>   * If...continually loading of slides while having
>> access to the reagent carousel is important to you, then I'd say Roche
>> Ultra has the advantage.
>>   *If...having more control of your IHC protocols to
>> change your retrieval conditions, or even loading one time retrieval
>> conditions without saving it as a new protocol, then I'd say Leica Bond has
>> the advantage.
>>
>> It really depends on what your lab will be staining is what I'm getting
>> to.
>> Another factor to include is ease of use.  Roche Ultra is literally a lot
>> easier to use than Leica Bond, and Leica Bond is easier to use than the old
>> Dako's or Dako Flex or similarly Thermo or Biocare IHC instruments.
>>* If you're staining odd protocols and want an
>> instrument staining biomarker assays at variant conditions outside the
>> norm:
>>  (ie: staining 50-100 micron frozen sections looking
>> for nerve bundles, or trying to perform FISH protocols on the instrument
>> without an installed FISH protocol on the instrument), then Leica is the
>> choice.
>>
>> quasi molecular staining:
>> 1.   if your lab is interested in using m-RNA
>> oligo probes, choose Leica, as they have an easier setup to using different
>> vendors other than their own mRNA detection.
>> 2.   if your physician staff prefers to see
>> probes with a matte background, only showing positive probe staining
>> (though using an older, yet very robust detection method), then Roche Ultra
>> has the advantage.
>>
>> If you're at a reference Lab and molecular performing testing that used
>> to be molecular based, but is now available via IHC, then Roche Ultra has a
>> huge advantage, particularly for the therapeutic and theranostic biomarkers
>> commercially available.
>>
>> Other factors to consider in your choice:
>> *Cost
>> * footprint of the instrument in your lab
>>* disposing of waste
>>* daily, weekly & monthly maintenance of the instrument
>>* cost of service.  Using a third party biomedical
>> group to repair or PM the instrument removes the IVD status of the
>> instrument, especially if you're staining any Class II's or higher, like
>> CD117, or ER, PR, Her2Neu...etc.
>> ***Its ok if you're just
>> using it for research if this doesn't apply.
>>
>> I hope this list helps you think what your team would like to use.  if
>> your pathologist and additional primary stakeholders would like to contact
>> me... you can.
>> I previously was a application specialist for IHC/probes with Leica
>> Microsystems up to 8 years ago.  However, I use a lot of Roche family of
>> instruments in my lab, as I'm a contracted lead for a well known
>> anatomic/clinical pathology laboratory.
>>
>> Best regards,
>> Eddie Martin, HT, HTL, QIHC
>>
>> On Thu, Oct 29, 2020 at 1:00 PM <
>> histonet-requ...@lists.utsouthwestern.edu> wrote:
>>
>>> Send Histonet mailing list submissions to
>>> histonet@lists.utsouthwestern.edu
>>>
>>> To subscribe or unsubscribe via the World Wide Web, visit
>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>> or, via email, send a message with subject or body 'help' to
>>> histonet-requ...@lists.utsouthwestern.edu
>>>
>>> You can reach the person managing the list at
>>> histonet-ow...@lists.utsouthwestern.edu
>>>
>>> When replying, please edit your Subject line so it is more specific
>>> than "Re: Contents of Histonet digest..."
>>> Today's Topics:
>>>
>>>1. Selection of IHC system (warda hassan)
>>>2. Re: Selection of IHC system (Tina Van Meter)
>>>
>>>
>>>
>>> -- Forwarded message --
>>> From: warda hassan 
>>> To: "his

Re: [Histonet] Histonet Digest, Vol 203, Issue 21

[Eddie Martin via Histonet]

Hi Histonet and W.A. Hassan,

Supposing you have a slide volume of 15-20k slides per year & currently use
a Dako instrument, but would like feedback on whether to get a:
  * Bond Max, or
  * Roche Ultra

My response would depend more than what your yearly slide volume is.
  * If...continually loading of slides while having access
to the reagent carousel is important to you, then I'd say Roche Ultra has
the advantage.
  *If...having more control of your IHC protocols to change
your retrieval conditions, or even loading one time retrieval conditions
without saving it as a new protocol, then I'd say Leica Bond has the
advantage.

It really depends on what your lab will be staining is what I'm getting
to.
Another factor to include is ease of use.  Roche Ultra is literally a lot
easier to use than Leica Bond, and Leica Bond is easier to use than the old
Dako's or Dako Flex or similarly Thermo or Biocare IHC instruments.
   * If you're staining odd protocols and want an
instrument staining biomarker assays at variant conditions outside the
norm:
 (ie: staining 50-100 micron frozen sections looking
for nerve bundles, or trying to perform FISH protocols on the instrument
without an installed FISH protocol on the instrument), then Leica is the
choice.

quasi molecular staining:
1.   if your lab is interested in using m-RNA
oligo probes, choose Leica, as they have an easier setup to using different
vendors other than their own mRNA detection.
2.   if your physician staff prefers to see
probes with a matte background, only showing positive probe staining
(though using an older, yet very robust detection method), then Roche Ultra
has the advantage.

If you're at a reference Lab and molecular performing testing that used to
be molecular based, but is now available via IHC, then Roche Ultra has a
huge advantage, particularly for the therapeutic and theranostic biomarkers
commercially available.

Other factors to consider in your choice:
*Cost
* footprint of the instrument in your lab
   * disposing of waste
   * daily, weekly & monthly maintenance of the instrument
   * cost of service.  Using a third party biomedical group
to repair or PM the instrument removes the IVD status of the instrument,
especially if you're staining any Class II's or higher, like CD117, or ER,
PR, Her2Neu...etc.
***Its ok if you're just
using it for research if this doesn't apply.

I hope this list helps you think what your team would like to use.  if your
pathologist and additional primary stakeholders would like to contact me...
you can.
I previously was a application specialist for IHC/probes with Leica
Microsystems up to 8 years ago.  However, I use a lot of Roche family of
instruments in my lab, as I'm a contracted lead for a well known
anatomic/clinical pathology laboratory.

Best regards,
Eddie Martin, HT, HTL, QIHC

On Thu, Oct 29, 2020 at 1:00 PM 
wrote:

> Send Histonet mailing list submissions to
> histonet@lists.utsouthwestern.edu
>
> To subscribe or unsubscribe via the World Wide Web, visit
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
> histonet-requ...@lists.utsouthwestern.edu
>
> You can reach the person managing the list at
> histonet-ow...@lists.utsouthwestern.edu
>
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
> Today's Topics:
>
>1. Selection of IHC system (warda hassan)
>2. Re: Selection of IHC system (Tina Van Meter)
>
>
>
> -- Forwarded message --
> From: warda hassan 
> To: "histonet@lists.utsouthwestern.edu"  >
> Cc:
> Bcc:
> Date: Wed, 28 Oct 2020 22:48:14 +0400
> Subject: [Histonet] Selection of IHC system
> Dear Histonet
>
> Our lab is looking to purchase a new IHC system.
> Currently we are working with Dako.
> I would request your valuable feedback on :-
>
> 1- Bond max ( Leica)
> 2- Bench Mark Ultra (Roche Diagnostic)
>
> Workload is 15,000-20,000 slides/year.
>
> Many thanks in advance for your help.
>
> Kind regards
> W.A Hassan
>
>
>
>
> -- Forwarded message --
> From: Tina Van Meter 
> To: warda hassan 
> Cc: Histonet@lists.utsouthwestern.edu
> Bcc:
> Date: Wed, 28 Oct 2020 23:08:07 -0400
> Subject: Re: [Histonet] Selection of IHC system
> Bond Max
>
> On Wed, Oct 28, 2020, 2:53 PM warda hassan via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
>
> > Dear Histonet
> >
> > Our lab is looking to purchase a new IHC system.
> > Currently we are working with Dako.
> > I would request your valuable feedback on :-
> >
> > 1- Bond max ( Leica)
> > 2- Bench Mark Ultra (Roche Diagnostic)
> >
> > Workload is 15,00

Re: [Histonet] Leica Bond, ASP6025 Processor parts

[Eddie Martin via Histonet]

Hi Paula, 

Are you using the Leica Bond for Clinical diagnostic testing? If yes, use of 
any biomed company to PM or use non-Original equipment manufacturing (OEM) 
parts on the instrument would void the IVD status of your instrument and any 
IVD reagents you are automating on it.  



Sent from my iPhone

>> On Aug 21, 2020, at 2:18 PM, Paula  wrote:
> Hi all,
> 
> 
> 
> Does anyone know of a Biomed company that makes and/or sells parts for the
> Bond III and the ASP6025 processor?
> 
> 
> 
> It's amazing how much Leica wants for their parts. For example, the waste
> bottle used to collect the buffer waste is $674 and the seal for the
> retort's lid is $597.  
> 
> 
> 
> Hope someone can help..thanks in advance.
> 
> 
> 
> Paula
> 
> Bio-Path Medical Group
> 
> Fountain Valley, CA

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[Histonet] Bubbles noticed on Bond-Max

[Eddie Martin via Histonet]

Hi Tim, 

From a user perspective: The slides you’re using may be a bad lot causing 
hydrophobic staining or you’re not cleaning your covertiles correctly. The 
wells on the assembly might also not be pulling adequately. Have you cleaned 
the assembly recently, particularly the wells? You could use a clean kit offer 
by Novocastra or make your own. Performing a decontamination on your instrument 
would be helpful too to include your bulk reagent compartments and reagent 
syringe.  Have you also been performing the suggested manufacturer’s guidance 
on maintaining your Bond-Max IHC/ISH analyzer?
Or...
Simply the instrument needs to be recalibrated by a Leica engineer. Do not PM 
with a third party biomed group as the Leica engineer will need to recalibrate 
the instrument with a PM kit to check and if necessary replace the physical 
components, electronics, and fluidic system of your analyzer along with 
updating the coordinate system to match software build according to their 
latest recommendation from their home office. 

Apart from typical lab reagents or processes only performed by an engineer,  do 
you have electrical or magnetic surges in your lab? The backup battery that 
comes with your Bond is typically good for two years, and would need to be 
replaced every few years, especially as the Bond-Max requires a step down from 
the 120V outlet to 110V. And this consideration can also deeply impact your 
overall staining. 


The application specialist should have more politely said that they didn’t know 
but will try to get a response back to you. I don’t comprehend how the tray 
would be the issue unless it was severely cracked and would prevent the slide 
assembly from functioning properly. 

Very respectfully,
Eddie Martin, HT, HTL, QIHC
Hematology Team Leader
The National Institutes of Health
10 Center Drive
Bethesda, MD 20892


Sent from my iPhone

> On Aug 13, 2020, at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote:
> 
> Send Histonet mailing list submissions to
>histonet@lists.utsouthwestern.edu
> 
> To subscribe or unsubscribe via the World Wide Web, visit
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
>histonet-requ...@lists.utsouthwestern.edu
> 
> You can reach the person managing the list at
>histonet-ow...@lists.utsouthwestern.edu
> 
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
> Today's Topics:
> 
>   1. Re: End of shift (Charles Riley)
>   2. Blank spots on IHC tissues - bubbles on Bond (Morken, Timothy)
>   3. Re: Blank spots on IHC tissues - bubbles on Bond (Morken, Timothy)
>   4. Re: Blank spots on IHC tissues - bubbles on Bond
>  (Paula Keene Pierce)
>   5. Protocol or SOP of how to make Tissue microarray
>  (Chakib Boussahmain)
> 
> 
> 
> 
> 
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Re: [Histonet] Quick question about Florida licensure.

[Eddie Martin via Histonet]

Hi Pam,

The quick response is that in Florida, an MLT with a MLT license cannot work as 
histotech unless they are working in Biomes, research, vending or in 
government. 

The Florida license specifies which type of clinical laboratories the 
technician/technologist is approved to work in. 

The Florida license states on it which laboratories the technician or 
technologist is approved to work in. 

If they request with the state to apply for state licensure in histology, then 
they must meet the requirements Prior to applying for licensure. 

V/r,
Eddie Martin


> On Aug 5, 2020, at 12:19 PM, Pam Barker  wrote:
> 
> Hi Histopeeps!
> I have a quick question about Florida licensure.  Can someone who holds a
> MLT(ASCP) get licensed and work as a histotech in Florida?
> 
> Thanks-Pam
> 
> Right Time, Right Place, Right Move with RELIA!
> *15 Years!*
> Celebrating 15 years of service exclusively to the Histology Community!
> 
> 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   
> check out our latest opportunities at:
> http://www.jobvertise.com/members/relia1 
> #jobs4myhistopeeps 
> #ilovemyhistopeeps
> #histopeeps
> Follow my hashtags and make your day great and your career greater!!
> 
> 
> 

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[Histonet] Recommend Harleco Giemsa stain reagents

[Eddie Martin via Histonet]

Milliliter sigma makes a brand called harleco for their giemsa’s line. I don’t 
think they make a kit. But you will need an 
1. neutral buffer reagent (harleco)
2.giemsa reagent
3. Azure B reagent 
And anhydrous methanol solution call me when you get these ans I can provide a 
working solution for your line up. 

V/r,
Eddie Martin
Hematology Team Leader 
NIH/CC/DLM Hematology Service
301-594-2054

Sent from my iPhone

> On Jul 27, 2020, at 10:46 AM, Katherine Bummer 
>  wrote:
> 
> Hello Histonetters,
> 
> Does anyone have a Giemsa stain they can recommend?  I prefer a kit and I 
> will be staining for neutrophils from a frozen cell pellet?
> 
> Thank you!
> 
> Kate
> 
> 
> This message is intended solely for the designated recipient(s). It may 
> contain confidential or proprietary information and may be subject to 
> attorney-client privilege or other confidentiality protections. If you are 
> not a designated recipient you may not review, copy or distribute this 
> message. If you receive this in error, please notify the sender by reply 
> e-mail and delete this message. Thank you.
> 

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Re: [Histonet] Can't keep a good histo tech down

[Eddie Martin via Histonet]

Hi Mike. I’m really sorry to hear that they’re terminating your role especially 
since you’ve filled that role for over 35 years at the same laboratory. 

FYI: Pam Barker with Relia is advertising a lead Ht role in a growing Tampa Bay 
lab. 

Pam’s number is 407-353-5070

I don’t want to just say “Best wishes in your new job search”. 

If you would like to talk, there are other histology contract providers I can 
send your way or discuss further with you. My office number will be posted 
below. I can schedule a break from work and discuss with you. 

For starters:
**Do you have a resume ready to go?
**Have you marketed yourself on job boards like indeed.com or monster.com? 
**Have you created a LinkedIn account to friend potential future jobs or job 
related connections that might provide an introduction to a position you would 
be intrigued to pursue? Or even list tasks you’re highly skilled at?
**do you have a digital trail of posts you replied to that show your subject 
matter expertise or willingness to help others that you could bring up during 
interviews?
**being a subject matter expert in histology is too great a skill set to keep 
in place. Have you considered working as a vendor for histology products or 
applications like IHC or selling equipment like microtomes and stainers?
***or considered going solo as a consultant or MOHS technician?

Your former job may have given you a dis-service...But it’s up to you on how 
you can make that turn around for yourself. This could be an opportunity to 
expand your wings into other histology related venues if you so wish. 


V/r,
Eddie Martin
NIH/CC/DLM Hematology Service
Team Leader
301-594-2054 office number

Sent from my iPhone

> On Jun 11, 2020, at 12:05 PM, MICHAEL S KATHY PENCE  
> wrote:
> 
> Yesterday I was called into my director's office and informed by the section 
> VP that they were eliminating my job of 35 years as Supervisor of Anatomic 
> Pathology. So here I am on Histonet seeing if anyone out there is looking for 
> a VERY skilled grossing HT for temp services. If you are feel free to contact 
> me and we can talk.
> 
> 
> 
> Sent from my U.S.Cellular© Smartphone
> 

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[Histonet] Xylene recycle or purchase fresh

[Eddie Martin via Histonet]

Hi Anne, 

If your xylene consumption is small, the footprint needed to recycle xylene in 
house and associated cost would be more expensive than buying xylene and 
shipping out the waste. 

V/r,
Eddie Martin



Sent from my iPhone

> On Jun 23, 2020, at 10:39 AM, Anne Murvosh  wrote:
> 
> I'm looking to switch from a disposal pick-up to recycling of Xylene. Can 
> anyone tell me some companies that offer a recycling unit. The catch is we 
> don't have very much so I need something for a small lab. Thanks Anne
> 

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Re: [Histonet] K and L

[Eddie Martin via Histonet]

Hi Melissa, 

It is possible to have a double negative K/L depending on patient’s diagnosis. 
It is also normal to have positive K/L expression in tissue samples. 

They are historically dirty in IHC. But you would have more control if you 
optimized with a concentrate version of the antibody instead of a Ready To Use. 
If you decide to still use the ready to use, you will have to add a lot of 
additional steps to reduce the background stain. 

Eddie Martin
Hematology & Histology Team Lead
The National Institutes Of Health
10 Center Dr
Bethesda, Maryland 20892


> On Jun 18, 2019, at 1:19 PM, Kuhnla, Melissa  wrote:
> 
> Good Afternoon,
> We are recently experiencing some staining issues with our IHC kappa and 
> lambda.  These antibodies have historically been problematic.  We are 
> currently running cell marque clones EP171 and EP 172 with Roche ultraview 
> detection.  The issues are complicated as sometimes they contradict each 
> other.  Lambda has too much background, which should be easy enough to 
> remedy.  Both stains also just recently have a lack of target specificity.  
> Only recently we have had multiple cases either both k and l are negative or 
> both stains are positive.  Anyone experiencing anything similar.  Thank you 
> in advance.
> 
> Melissa Kuhnla
> Lead Medical Technologist
> for IHC and FISH testing
> Regional Laboratory Services
> Good Samaritan Hospital
> 70 Arkay Dr.
> Hauppauge, NY 11788
> 631-609-2551
> 
> The information in this e-mail, and any attachments therein, is confidential 
> and for use by the intended addressee only. If this message is received by 
> you in error please do not disseminate or read further. Please reply to the 
> sender that you have received the message in error, then delete the message. 
> Although Catholic Health Services of Long Island attempts to sweep e-mail and 
> attachments for viruses, it does not guarantee that either are virus-free and 
> accepts no liability for any damage sustained as a result of viruses. Thank 
> you.
> 

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Re: [Histonet] CC1 vs Ultra CC1

[Eddie Martin via Histonet]

Hi Curt,

It technically would work. However it wouldn’t be GLP to do so (for several 
reasons). 

However, if you’re using the Ultra for research, non-Diagnostic purposes,  and 
you were still hesitant to mix cc1 with ultra cc1, then you could Decon CC1 and 
fill with the XT CC1. When you receive the Ultra CC1, Decon again to remove the 
XT CC1 and fill with the Ultra CC1. 

Hope this helps,
Eddie

Sent from my iPhone

> On Jun 3, 2019, at 3:15 PM, Curt Tague  wrote:
> 
> I made the mistake of ordering CC1 instead of Ultra CC1 I guess that 
> can't be used on the benchmark ultra It's an "ULTRA" thing I guess...
> 
> Anyway, anyone out there use regular CC1... want a couple bottles? Brand new, 
> unopened...
> 
> Curt
> 
> 

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Re: [Histonet] IDH1

[Eddie Martin via Histonet]

Hi Richard,

We used Ventana H09 clone for IDH1 while I worked at Walter Reed NMMC. 

Sent from my iPhone

> On Dec 14, 2018, at 3:38 PM, Cartun, Richard  
> wrote:
> 
> For those labs that are performing IDH1 IHC testing on FFPE tissue, which 
> clone are you using and where do you get it?  Thank you.
> 
> Happy Holidays to everyone.
> 
> 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 reply e-mail and destroy all copies of the original 
> message, including any attachments.
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Re: [Histonet] BMP-2 protocol for bone

[Eddie Martin via Histonet]

Hi Lindsay, 

There are a few complications  that will automatically come up due to working 
with rabbit tissue, and also due to the antibody from Abcam that you are using. 

I called Abcam prior to emailing you on a hunch that I had and was correct. The 
antibody you purchased from Abcam is unconjugated and first needs to be 
conjugated to whatever method you’re going to use. You could conjugate the 
antibody yourself. I already reached out to Abcam. It’s going to take them a 
few business days to get back to me to see whether their own antibody can be 
conjugated for IHC if that’s the route you’re going to go. 

Keeping this email simple for a response to your question, it may be easier to 
use another detection method for conjugating this specific antibody to rabbit 
bone...possibly FITC conjugated tyramide labeling with Alexa-Fluor. 

If you are insistent on using HRP it may be easier to contact me and we can 
further discuss as there are easily two complications that will arise due to 
the antibody you chose to optimize with and the species you are trying to stain 
on. 

Since you left your lab number on your post, I called and left my phone number 
on the voicemail for the young Lab if you would like to discuss and 
troubleshoot over the phone. I can also be reached via email at work at 
eddie.mar...@nih.gov

Thanks,
Eddie

Eddie Martin HT(ASCP), QIHC(ASCP),HTL
Hematology Team Lead / Histology Technical Supervisor
The National Institutes of Health 
Clinic Center - Department of Laboratory Medicine
10 Center Drive, Room 2C 360
Bethesda, MD 20814



> On Oct 18, 2018, at 12:57 PM, Wilson, Lindsay A 
>  wrote:
> 
> Good morning,
> 
> Does anyone have a proven IHC protocol for BMP-2 on bone tissue? A big plus, 
> if you have worked with it on rabbit bone!
> I am currently using Abcam’s BMP-2 (ab6285) with no success. I have performed 
> antigen retrieval and non-antigen retrieval. I have used a dilution as low as 
> 1:50. I appreciate any suggestions.
> 
> Lindsay Wilson, LVT, RLATG
> Young Laboratory
> 
> UTHealth | The University of Texas Health Science Center at Houston | School 
> of Dentistry
> Department of Oral & Maxillofacial Surgery
> 
> 7500 Cambridge St. | Suite 6510 | Houston, TX 77054
> 713 486 4360 tel | 713 486 4333 fax
> 
> 

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Re: [Histonet] Restaining old Heamatology smears

[Eddie Martin via Histonet]

Hi Laurie,

Blood is considered a tissue so it is very much still Histo related. 

Regarding your smears, this also occurs in our hematopathology laboratory from 
time to time when we are scanning old slides and there’s a bubble under the 
coverslip that won’t let us take images with our digital scope. 

It may very well be that there is still a resinous permount film on your slides 
from the old coverslip. This may simply be removed by leaving the smear in 
xylene some additional time to remove the resin. Then place slides in 100% 
alcohol to remove xylene residues. The following steps would be rehydrating the 
slides with dilute changes in in alcohol...ie: 95%,then 85%, then 70%, then in 
a few changes of distilled water. If you can’t get the pH of your water to pH 
7.2, then you could use a phosphate buffered solution to achieve this in place 
of the distilled water. 

Then place your slides in a working Giemsa working  solution. It may help to 
rock your slides or slightly agitate them to improve staining intensity. 

I hope this helps. 
Please feel free to contact me via email at eddie.mar...@nih.gov





> On Oct 17, 2018, at 10:51 PM, Reilly, Laurie  wrote:
> 
> Dear Histonetters,
> This is not quite Histo but related. One of our Laboratory Scientists has 
> removed the coverslips from faded blood smears used for teaching and is 
> having trouble restaining them with Wright's stain.
> Could we please call on the combined wisdom of the Histonet to try to find a 
> solution to this problem.
> 
> Regards,  Laurie.
> 
> Mr. Laurie REILLY
> Histopathology
> Veterinary and Biomedical Sciences
> James Cook University
> Townsville  Qld.  4811
> Australia.
> 
> Phone 07 4781 4468
> Mobile 0448 957747
> 
> 

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[Histonet] Prostate block chipping out

[Eddie Martin via Histonet]

Hi Mary Ann,

It seems that you’re having a problem with either your block assembly or your 
blade holder assembly.  I would have your microtome serviced, it may be the 
block clamp isn’t tight enough or you have knicks on your blade holder.  If 
you’re block clamp is ok, I would lean more on the blade holder needing to be 
repaired or replaced. The older blade holder assemblies cost more, as they were 
fabricated to last longer before giving issues with microtomy.  The newer 
microtome assemblies can easily knick and give thick and thin sections, and in 
this case giving 1 good section, followed by a section that is chunking out.

Hope this helps!

Cheers,
Eddie Martin, HTL, HT(ASCP), QIHC(ASCP)
Histology Technical Supervisor
The National Institutes of Health
The Clinic Center
Department of Laboratory Medicine
10 Clinic Dr
Bethesda, MD 20814

> On Jul 8, 2018, at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote:
> 
> Send Histonet mailing list submissions to
>   histonet@lists.utsouthwestern.edu
> 
> To subscribe or unsubscribe via the World Wide Web, visit
>   http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
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> 
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
> Today's Topics:
> 
>   1. Prostate biopsy (Mary Ann)
> 
> From: Mary Ann 
> Subject: [Histonet] Prostate biopsy
> Date: July 7, 2018 at 2:29:13 PM EDT
> To: histonet@lists.utsouthwestern.edu
> 
> 
> I have an issue with our prostate needle biopsy chipping out of the block on 
> sectioning.
> One block out of 12 will demonstrate this phenomenon.  The block will section 
> well then a section with chip right out.
> 
> Sent from my iPhone
> 
> 
> 
> 
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[Histonet] Leica ST5050 immunostainer

[Eddie Martin via Histonet]

Dr Fleming,

If you’re insistent on automation, I would suggest a used Leica Bond Max 
instead.  The Leica Bond Max is most likely the most cost effective, and has 
the smallest footprint while also minimizing reagent waste compared to a Dako 
Flex or Ventana XT/Ultra. 

If you are willing to do it by hand, there a few labs that I’m aware of that 
use Leica’s powervision  compact polymer system to stain their Melan-A’s. I’m 
sure it would stain a MITF just as easy. 

Best,
Eddie M. 

Team lead medical technologist
Histology Technical Supervisor
The National Institutes of Health 
Clinic Center
Bethesda, MD 20814
eddie.mar...@nih.gov



> On Mar 16, 2018, at 9:00 AM, Matthew Fleming  wrote:
> 
> Hi,
> 
> I have a small dermatopathology lab and would like to introduce limited
> immunohistochemistry.  Initially I would probably just do MITF stains with
> red chromogen on a few cases per week. We could do this by hand but I've
> noticed a number of ST5050 immunostainers for sale at very low prices. I
> know they're quite old but we have a number of machines old or older that
> continue to serve us well. But so far I haven't been able to find a manual
> and am concerned about finding consumables. Leica apparently stopped
> supporting this model some time ago.  Is anyone out there still using it?
> Anyone have a manual? Anyone like to comment as to whether it might be
> usable for a limited IH operation?
> 
> Thank you,
> 
> Matthew Fleming, MD
> Fleming Dermatopathology
> Milwaukee, WI
> 

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[Histonet] validation inquiry regarding negative tissue

[Eddie Martin via Histonet]

Hi Nancy,

The use of negative internal control tissue on your antibody testing for 
negative cases would have to be determined by your medical director/chief 
pathologist as acceptable. 

You may also use Lipoma tissue as a negative control tissue. It works as a 
negative control for just about every antibody I can think about. There may 
also be a few antibodies that would have false positive staining with a lipoma, 
but a pathologist could read through that stuff. 

Hope this helps!

Kind regards,
Eddie M.

Edward M. Martin, HTL, HT(ASCP), QIHC(ASCP)
Lead Hematopathology Medical Technologist
The National Institutes of Health
and
IHC Histotechnologist III
The Joint Pathology Center
Walter Reed NMMC
Bethesda, MD


> On Oct 24, 2017, at 1:16 PM, Nancy Schmitt  wrote:
> 
> Good Day!
> When performing validation on IHC, we use 20 positives and 20 negative cases.
> 
> When testing for negative, can the normal tissue surrounding the tumor be 
> counted as negative?
> 
> Does the negative tissue need to be tumor that is negative for the AB or just 
> normal tissue?
> 
> Thank you for your consideration,
> 
> Nancy
> Dubuque, IA
> Ph. 563-589-9810
> 
> 
> 
> NOTICE: This email may contain legally privileged information. The information
> is for the use of only the intended recipient(s) even if addressed
> incorrectly. If you are not the intended recipient, please notify the sender
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> 
> 
> 
> 
> NOTICE: This email may contain legally privileged information. The information
> is for the use of only the intended recipient(s) even if addressed
> incorrectly. If you are not the intended recipient, please notify the sender
> that you have received it in error and then delete it along with any
> attachments. Thank you.
> 
> 
> 

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Re: [Histonet] Running ISH on the Ventana Ultra Stainers

[Eddie Martin via Histonet]

Hi Miranda,

For starters, perform a Decontamination on your Ultras, don't dilute your 
probes. At our lab, we manually dispense the probes but a prep kit is fine.  
And also use a toothpick to clear out any gunk in your detection dispensers 
prior to doing a run. Also, check if your ultras are on battery backups 
providing sufficient run time amperage in case of power failures or 
abnormalities. Also moving your ultras or the instrument not being level can 
create issues. And check that you lo of charged slides are staining well across 
several antibodies. 

If all that is ok, then try out your protocol again and see if you get adequate 
staining quality. 

Hope this helps to get you started!

Eddie Martin, HT,HTL,QIHC
IHC Histotechnologist III
The Joint Pathology Center
Walter Reed National Military Medical Center
Bethesda, MD

Sent from my iPhone

> On Sep 19, 2017, at 1:25 PM, Miranda Giorgi  wrote:
> 
> Hello Histonetters,
> 
> Our lab is in the process of trying to validate Kappa, Lambda and EBER ISH on 
> the Ventana Ultra stainers.  We have run these probes on the XT in the past 
> with good results, but unfortunately staining on the Ultras is either poor or 
> non-existent.  We have tried several protocol modifications with very little 
> luck.  Does anybody out there have experience validating these tests on the 
> Ultras?  We would greatly appreciate any advice on getting these to work.
> 
> Thanks in advance!
> 
> 
> This e-mail and any attachments may contain CONFIDENTIAL information, 
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> recipient, any use or disclosure of this information is STRICTLY PROHIBITED; 
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[Histonet] PIN4 not working

[Eddie Martin via Histonet]

Hi Kim,

I was wondering if you could be more descriptive as to what is going wrong with 
your PIN4 stain?

It's difficult to give a suggestion when we don't have any clue as to what your 
lab is visualizing. 

Is it the DAB?
Is it the AP Fast Red chromagen?
Is it reduced staining or no staining?

Kind regards,

Eddie Martin, HTL, HT(ASCP), QIHC(ASCP)
IHC Histotechnologist III
Joint Pathology Center (JPC)
Walter Reed National Military Medical Center
Bethesda, MD
 



> On Aug 4, 2017, at 5:06 PM, Renee Fisher  wrote:
> 
> Need help the PIN4 has suddenly stop working we use P63+CK5+CK14 from Bio 
> Care Medical and P504S from Dako.  We double stain on the Leica Bond with the 
> Polymer refine Red and Polymer Refine detection. This stain has worked 
> beautifully in the past. Any suggestions?
> 
> 
> 
> Thank you
> 
> 

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Re: [Histonet] CPT Code for PGP9.5 free floating tissue stain

[Eddie Martin via Histonet]

You can only bill the 88342 once for the initial IHC with interpretation charge 
for the pgp9.5, (assuming it was the first antibody you billed to the patient). 
 The pgp9.5 is the test your pathologist ordered. The linker step in your 
protocol isn't another test ordered, just part of your validated protocol in 
order to make the pgp9.5 work correctly. 

If your pathologist ordered another antibody test on the same specimen part 
other than pgp9.5, then you could apply the 88341 cpt code. For example, 
ordering an s-100p on the same specimen would be an 88341 on the same specimen 
part. 

However, if you order another pgp9.5 on the same specimen part, then it would 
be billed as a no charge.

Hope this helps. 





Sent from my iPhone

> On Jun 22, 2017, at 3:21 PM, Haley Huggins  wrote:
> 
> I am running into an issue with billing and which code is correct for
> staining PGP9.5. Is it 88342? Also the billing guy asked me if thr
> secondary goat, anti-mouse secondary antibody could be billed, can it? I
> would appreciate any help I can get as soon as possible! He needs to bill
> out today. Thanks.
> 

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[Histonet] Response Fixation and IHC for animal tissue

[Eddie Martin via Histonet]

Hi,

CAP guidelines don't cover animal testing or research testing. 

V/r,

Eddie Martin HT, HTL, QIHC

Sent from my iPhone

> On Jun 14, 2017, at 7:15 PM, Cristi Rigazio  wrote:
> 
> Hi histonetters!
> 
> Can anyone tell me what, if any, guidelines there are for fixation time for
> animal tissue with potential subsequent IHC stains.  I am very familiar
> with CAP guidelines for receptor testing with breast cases, but I can't
> seem to find much on IHC for animal tissue.
> 
> Thanks,
> Cristi
> 

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Re: [Histonet] What happened to Ultraclone/PGP9.5?

[Eddie Martin via Histonet]

Novocastra has been selling pgp 9.5 for years. I've used it on frozen sections 
and FFPR on 3 microns as well as 50 micron sections and it stains very well. 

Sent from my iPhone

> On Jun 3, 2017, at 3:22 PM, Hobbs, Carl  wrote:
> 
> 
> 
> Ultraclone
> THE supplier of PGP9.5 for many years !
> Respect to them/him/her/LTB
> No longersigh.
> Ultraclone:  "legendary"...chuckle
> 
> 
> Sureother Suppliers now offer anti PGP9.5 abs
> Why then do they still call it "PGP9.5"??
> We now know what the protein is.
> 
> PGP: "Pretty good protein" I recall?
> Why the 9.5?
> Usually a clone but, but Ultraclone ab was a rabbit poly.
> 
> One could only order via fax, I recall.
> Isle of Wight location: Rossiters Farm.
> 
> Any more  info re Ultraclone's history  would be most appreciated.
> 
> Curiously
> 
> Carl  
> 
> 
> 

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Re: [Histonet] CAP checklist help

[Eddie Martin via Histonet]

Hi Charles,

The Leica BOND system displays the temperatures of the individual heating pads 
during the retrieval stage on your computer monitor. You can easily take a 
screenshot of your computer monitor and save it to a flash drive as a digital 
record.
*A heating error would appear if the individual heating pad didn't reach 100 
degrees. 

 The Leica service engineer for the Bond can also provide documentation every 
time they service your IHC analyzer. You can also check your slide staining 
history and find cases that didn't stain well and individually check if the 
result was due to the temperature not reaching 100 degrees on that individual 
heating pad. Hope this helps. 

V/r,
Eddie Martin, HTL, QIHC
IHC Histotechnologist III
Walter Reed National Military Medical Center 


> On Dec 28, 2016, at 11:55 AM, Charles Riley  wrote:
> 
> I need help with figuring out how to meet checklist # ANP.23420.   We use
> the Leica Bond III and MAX  to do our ISH slides.  How do I test the
> temperature and how would you recommend recording the results?
> 
> -- 
> 
> Charles Riley HT(ASCP)CM
> 
> Histopathology Coordinator/ Mohs
> 

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Re: [Histonet] Low volume IHC instrument

[Eddie Martin via Histonet]

Hi Gareth, 

If Fako doesn't suit your needs, then bio are wouldn't either. The closest 
analyzer with full onboard retrieval that is also more open without purchasing 
separate detection kits to improve quality is the Leica Bond. H pylori 
retrieval on Bond is only 10 mins and I think CD8 is 20 mins I think. So the 
overall stain time would be a few hours on the Leica Bond-max and about an hour 
less on the Bond-III. It shaves off at least 45 mins from the time spent by a 
bond-max platform. 

I have also used Ventana XT, and ultra. We can discuss further if you would 
like to call me at 954-826-9403. 

Hope this helps,
Eddie Martin HT(ASCP),QIHC, HTL

Sent from my iPhone

> On Sep 15, 2016, at 5:37 PM, Gareth Davis  wrote:
> 
> What IHC platform would you choose if you only stained for CD8 and H.
> pylori?
> Ventana, Leica Bond, or Biocare?  Or is there another I am missing?
> Familiar with Dako, and don't think it would suit our lab.
> Thanks again.
> 
> 
> -- 
> Ms. Gareth B. Davis, HT, QIHC (ASCPcm)
> Yuma Gastroenterology
> Yuma, AZ 85364
> 928-248-5259
> 

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Re: [Histonet] slides on the Benchmark Ultra

[Eddie Martin via Histonet]

Hi Amy,

While I worked at the University of Alabama in Birmingham, they used the super 
frost plus slides provided through Cardinal Health. They were the same Fisher 
slides to be used on the Ventana IHC analyzers. I hope this helps. 

Best,
Eddie Martin, HT(ASCP),QIHC,HTL

Sent from my iPhone

> On Aug 3, 2016, at 9:06 AM, Amy Johnson  wrote:
> 
> Hello Histonetters,
> 
> WE currently run the Benchmark Ultra for our IHC staining.  We have been 
> using SuperFrost Plus slides on the recommendation by Ventana.  What are 
> others using if you aren't using SuperFrost Plus slides.
> Our vendor that we get SuperFrost Plus from no longer carries them and has 
> replaced them with another type..Just wondering what others think
> 
> 
> Thanks
> Amylin Johnson, B.S. HTL(ASCP)
> Associates in Pathology
> Wausau Wi 54401
> 715-847-2130
> 
> 

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Re: [Histonet] MITF

[Eddie Martin via Histonet]

Hi Kathy,

The Journal of Investigative Dermatology had an article in their December 2011 
Volume 131 issue 12 on Human Cutaneous Melanomas lacking MITF and melanocyte 
differentiation. According to the article, Melanomas could be MITF and Melan-A 
negative, if the Melanomas were TAM tyrosine kinase receptor positive. In this 
particular study, 38% of the total Cutaneous Melanomas were Axl+ (member of 
TAM), and none of the Axl+ were MITF and Melan-A positive.

That being said, it may explain why some of your cutaneous melanomas are not 
MITF positive.  Known internal control tissue for MITF includes epithelioid 
tissue, mast cells, reactive histiocytes and osteoclasts stain intensely for 
MITF.  A nevus would also be an ideal control tissue, as would any other 
perivascular epithelioid tumor.

Best,
Eddie Martin, HTL, HT(ASCP), QIHC
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[Histonet] Why is Auromine heated in Histology, but not in Microbiology

[Eddie Martin via Histonet]

Hi Angie, 

The simplest answer, would be to save time.  Whether heated, or microwaved at 
80 power for 45 seconds, would be quicker than flooding the slide and letting 
stand for extended periods of time.
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Re: [Histonet] CD34 for primate tissue

[Eddie Martin via Histonet]

Hi Cathy,

I haven't heard of the cross reactivity with monkey species using Novocastra 
CD34. However, novocastra's antibody is a monoclonal antibody. You may get 
better staining utilizing a polyclonal CD34 instead. Thermo-Fisher and Abcam 
provide a polyclonal CD34. If you reach out to them, they may be able to 
provide you with a sample size 0.1 microliter if they have them available. 

I hope his helps!

Best,
Eddie


Sent from my iPhone

> On Jun 30, 2016, at 9:06 AM, Cathy M. Mathis/Comparative Medicine 
>  wrote:
> 
> Rhesus is a species of old world monkey.  There are a couple of other 
> companies that have this same clone and on their datasheets they say it cross 
> reacts with rhesus monkey.  The CD34s (2) that I have do not state on their 
> data sheets that they cross react with anything except human and yet are the 
> same clone as the companies that say it does.  I guess I will just break down 
> and purchase a different clone that states it works in rhesus monkey.
> Cathy
> 
> -Original Message-
> From: Eddie Martin [mailto:edmarti...@gmail.com] 
> Sent: Wednesday, June 29, 2016 8:22 PM
> To: Cathy M. Mathis/Comparative Medicine
> Cc: histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] CD34 for primate tissue
> 
> What species is the kidney you are trying to stain with IHC. Because you 
> mentioned that you used normal human kidney and it worked , but Rhesus kidney 
> didn't stain. I'm wondering if the Rhesus kidney is human or another animal 
> species? The Novocastra CD34 (QBEND10) is intended for human tissue. 
> 
> Best,
> Eddie Martin, HTL,QIHC
> 
> Sent from my iPhone
> 
>> On Jun 29, 2016, at 4:43 PM, Cathy M. Mathis/Comparative Medicine 
>>  wrote:
>> 
>> 2 more bits of information about my dilemma; I did try staining 
>> without any retrieval - no signal I am using rhesus kidney as a 
>> control (getting no signal), but I also ran some human kidney and got 
>> beautiful staining with a 20 minute HIER using a pH 8 EDTA buffer.
>> So I know the antibodies and my protocols work, just not on my rhesus.  Does 
>> anyone know of a CD34 that will work in this species?
>> More suggestions please?  Thank you all for your time.
>> Cathy
>> 
>> -Original Message-
>> From: Eddie Martin [mailto:edmarti...@gmail.com]
>> Sent: Wednesday, June 29, 2016 5:34 PM
>> To: Cathy M. Mathis/Comparative Medicine
>> Cc: histonet@lists.utsouthwestern.edu
>> Subject: Re: [Histonet] CD34 for primate tissue
>> 
>> Hi Cathy,
>> 
>> EDTA (Retrieval #2) for 20 minutes on the Bond-Rx should be sufficient. And 
>> all that is necessary. Please contact me if you need additional help. 
>> 
>> Best,
>> Eddie Martin, HTL, QIHC
>> edmarti...@gmail.com
>> 
>> Sent from my iPhone
>> 
>>> On Jun 29, 2016, at 8:16 AM, Cathy M. Mathis/Comparative Medicine 
>>>  wrote:
>>> 
>>> Good morning my fellow Histo-netters, Does anyone have experience 
>>> with any CD34 antibody that would work in FFPE rhesus tissues?  A few 
>>> companies have one with clone QEBend 10 and say that it works but I have 
>>> had no luck.  I am using the Bond RX polymer system and I've tried all the 
>>> epitope retrievals from Leica.  Now I am try epitope retrieval offline; 
>>> pronase, trypsin, pressure cooker with high and low pH solutions.  Still no 
>>> signal.  Any help would be greatly appreciated.
>>> Cathy
>> 
> 
> 
> 

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Re: [Histonet] CD34 for primate tissue

[Eddie Martin via Histonet]

What species is the kidney you are trying to stain with IHC. Because you 
mentioned that you used normal human kidney and it worked , but Rhesus kidney 
didn't stain. I'm wondering if the Rhesus kidney is human or another animal 
species? The Novocastra CD34 (QBEND10) is intended for human tissue. 

Best,
Eddie Martin, HTL,QIHC

Sent from my iPhone

> On Jun 29, 2016, at 4:43 PM, Cathy M. Mathis/Comparative Medicine 
>  wrote:
> 
> 2 more bits of information about my dilemma;
> I did try staining without any retrieval - no signal
> I am using rhesus kidney as a control (getting no signal), but I also ran 
> some human kidney and got beautiful staining with a 20 minute HIER using a pH 
> 8 EDTA buffer.
> So I know the antibodies and my protocols work, just not on my rhesus.  Does 
> anyone know of a CD34 that will work in this species?
> More suggestions please?  Thank you all for your time.
> Cathy
> 
> -Original Message-
> From: Eddie Martin [mailto:edmarti...@gmail.com] 
> Sent: Wednesday, June 29, 2016 5:34 PM
> To: Cathy M. Mathis/Comparative Medicine
> Cc: histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] CD34 for primate tissue
> 
> Hi Cathy,
> 
> EDTA (Retrieval #2) for 20 minutes on the Bond-Rx should be sufficient. And 
> all that is necessary. Please contact me if you need additional help. 
> 
> Best,
> Eddie Martin, HTL, QIHC
> edmarti...@gmail.com
> 
> Sent from my iPhone
> 
>> On Jun 29, 2016, at 8:16 AM, Cathy M. Mathis/Comparative Medicine 
>>  wrote:
>> 
>> Good morning my fellow Histo-netters,
>> Does anyone have experience with any CD34 antibody that would work in FFPE 
>> rhesus tissues?  A few companies have one with clone QEBend 10 and say that 
>> it works but I have had no luck.  I am using the Bond RX polymer system and 
>> I've tried all the epitope retrievals from Leica.  Now I am try epitope 
>> retrieval offline; pronase, trypsin, pressure cooker with high and low pH 
>> solutions.  Still no signal.  Any help would be greatly appreciated.
>> Cathy
> 

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Re: [Histonet] CD34 for primate tissue

[Eddie Martin via Histonet]

Hi Cathy,

EDTA (Retrieval #2) for 20 minutes on the Bond-Rx should be sufficient. And all 
that is necessary. Please contact me if you need additional help. 

Best,
Eddie Martin, HTL, QIHC
edmarti...@gmail.com

Sent from my iPhone

> On Jun 29, 2016, at 8:16 AM, Cathy M. Mathis/Comparative Medicine 
>  wrote:
> 
> Good morning my fellow Histo-netters,
> Does anyone have experience with any CD34 antibody that would work in FFPE 
> rhesus tissues?  A few companies have one with clone QEBend 10 and say that 
> it works but I have had no luck.  I am using the Bond RX polymer system and 
> I've tried all the epitope retrievals from Leica.  Now I am try epitope 
> retrieval offline; pronase, trypsin, pressure cooker with high and low pH 
> solutions.  Still no signal.  Any help would be greatly appreciated.
> Cathy
> 
> 
> 

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Re: [Histonet] Factor VIII

[Eddie Martin via Histonet]

Hi Carol,

You probably are getting inconsistent results with Novocastra Factor 8 antibody 
because the Novocastra RTU is for human diagnostic testing. If you need it for 
canine testing and would still need to use the bond max to perform the test, 
then you would need a canine anti-mouse or Canine anti-rabbit preferably 
monoclonal antibody. 

Kind Regards,
Eddie Martin HT(ASCP),QIHC
954-826-9403

Sent from my iPhone

> On May 25, 2016, at 2:58 PM, Johnson, Carole  wrote:
> 
> Hello all,
> I have been working up the Factor VIII (RTU) on a Bond Max platform with 
> limited success on canine tissue.  Does anyone have a favorite that they can 
> recommend?
> 
> Thanks in advance,
> Carole
> 
> Carole L. Johnson, HT(ASCP)cm, QIHC
> 
> New Mexico Department of Agriculture
> Veterinary Diagnositc Services
> 1101 Camino de Salud, NE
> Albuquerque, NM 87101
> 505.383.9299
> 
> 
> 
> 
> Confidentiality Notice: New Mexico has a very broad public records law. Most 
> written communications to or from state employees are public records. Your 
> e-mail communications may therefore be subject to public disclosure. This 
> e-mail, including all attachments is for the sole use of the intended 
> recipients. Any unauthorized review, use, disclosure or distribution is 
> prohibited unless specifically provided under the New Mexico Inspection of 
> Public Records Act.
> 

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[Histonet] Tips on staining PLA2R assay on Leica Bond

[Eddie Martin via Histonet]

I don’t do staining for this antibody, but have experience with IMF on frozen 
tissue and FFPE and have worked with the Leica BOND max and BOND-3.  If you 
would prefer IHC over IMF, you can search for a human anti-mouse, or human 
anti-rabbit antibody.  Abcam provides a polyclonal whole serum antibody that 
works on FFPE.  I grabbed this from their website: 
Anti-PLA2R antibody (ab80054)

Since Abcam’s PLA2R antibody is a rabbit anti-human, you would need to create a 
modified DAB or modified RED protocol that doesn’t include the secondary 
antibody / linker step in your protocol prior to dispensing polymer.  The rest 
of your protocol can remain as it is.  

As you are using a Leica BOND, both ways are actually pretty easy to set up, 
either indirect IMF or using Leica’s DAB kit for IHC. You would just need to 
play around with retrieval times with both Citrate and EDTA to get the optimum 
staining pattern. Additional ancillaries may be added when necessary should you 
have any background stain that is hard to get out.

Best Regards,

Eddie Martin, HT(ASCP), QIHC
954-826-9403
edmarti...@gmail.com
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Re: [Histonet] Automated IHC instrument

[Eddie Martin via Histonet]

Hi Valerie,

Any platform would get the job done well.  They all offer reagent rental 
agreements too.  Dako and Leica would require the user to be a bit more 
knowledgeable of IHC protocols than starting a run on a Ventana analyzer.  Dako 
is semi-automated, whereas leica and ventana both have onboard retrieval, and 
reagent mixing of chromagen.  In terms of the most open system, Dako would be 
the best analyzer followed by Leica in second place.  Ventana does allow 3rd 
party antibodies to be run on it, but charge you a premium for open containers 
to run on their analyzer.

In terms of antibodies: this may be better chosen by your pathologist which may 
have an impact on which analyzer you will also get, but I can suggest to you my 
favorites.

Her2neu - all three vendors provide a good Her2neu clone. Dako’s Her2 sometimes 
doesn’t stain as well, so would be my least favorite of the 3. Ventana’s Her2 
is the most widely used.

ER - all 3 vendors are also good.

PR - Dako and Novocastra have great PR clones.  Ventana’s PR clone is IVD as 
Dako and Novocastra Clone but is not ASCO-CAP approved antibody.  Ventana will 
push to say it is IVD, as Dako, and Novocastra (Leica), which is necessary for 
billing purposes, but their PR clone isn’t as effective as the other two i 
mentioned.

Ki67 - Dako’s MIB-1 clone is like the gold standard for Ki-67.  All three 
vendor versions work as intended.  Leica removed one of their two clone options 
last year, so k2 clone for Leica is your only option.  Ventana’s clone looks 
nicer in my opinion.

Hope this helps somehow.

Kind Regards,

Eddie Martin, HT(ASCP), QIHC, HTL
954-826-9403
edmarti...@gmail.com 
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