[Histonet] Leica Bond Max artifact
Cathy M. Mathis/Comparative Medicine
cmmathis at wakehealth.edu
Wed Oct 10 12:41:02 CDT 2018
Hello Dr. Rosen and Histonetters,
We have used a Leica Bond RX for over 2 years now, cleaning and caring for it just as you do. My work is mostly animal which is why we mainly choose the red chromogen alkaline phosphatase detection kit. We see the same artifact sporadically. Sometimes we will see it for several days and then maybe a week goes by and we don't see any of it. It may only be a small amount and I have seen it basically all over the slide. I know it is not related to our tissues or slides. Our Leica rep came out and increased washes but it really doesn't help. If anyone has a solution to the problem, I would also be interested.
Best to all,
Cathy
Cathy M. Mathis
Laboratory Technician IV
Department of Pathology
Section on Comparative Medicine - Clarkson Campus
Medical Center Boulevard \ Winston-Salem, NC 27157
p 336.716.1538 \ f 336.716.1515
cmmathis at wakehealth.edu wakehealth.edu
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Sent: Wednesday, October 10, 2018 1:00 PM
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Subject: Histonet Digest, Vol 179, Issue 9
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Today's Topics:
1. FW: Leica Bond Max artifact (Linda Margraf)
2. Re: FW: Leica Bond Max artifact (Rene J Buesa)
3. H&E for H. pylori? (P Sicurello)
4. Re: H&E for H. pylori? (Rene J Buesa)
5. Re: H&E for H. pylori? (Tony Henwood (SCHN))
6. Job Opening in Massachusetts: IHC Lab Supervisor (Erin Sarricks)
7. Low Oxygen Sensor/Alarms (Cooper, Brian)
----------------------------------------------------------------------
Message: 1
Date: Tue, 9 Oct 2018 17:06:27 +0000
From: Linda Margraf <Linda.Margraf at cookchildrens.org>
To: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] FW: Leica Bond Max artifact
Message-ID: <d335fdb52e5b4e93bfb17bad8554cb02 at MBX10.CCHCS.LDAP>
Content-Type: text/plain; charset="utf-8"
Here is a message I am posting for Dr. Rosen:
Hello,
I am a longtime admirer of this list, and I am sure someone on here can help us out
We have an artifact on our immunohistochemistry (IHC) slides.
I will attempt to attach pictures using the upload tool. In case that doesn?t work, I am attaching a google drive link to some jpgs. The artifact is usually golden brown to red/pink in color, is clustered, appears both on top of the tissue and around it. If we do an aggressive rundown, it appears less red/brown and bluer.
We are using a Leica Bond MAX IHC system which is about a year old. We usually do two runs a day and 95% of the time we only use a red detection kit. We only see this on our red detection kit on our Leica bond. The brown is unaffected. The corresponding H&E slides are fine.
We started having this artifact after a month or two of production. 85% of the time we have some sort of artifact. Sometimes its perfect, sometimes it makes the tissue unreadable. We haven?t figured out any precipitating factors.
We have tried a number of things, but we still can?t get control of this.
Here is a brief overview of our protocols:
- Processor: Leica ASP300s tissue processor.
-Parrafin = Mercedes HWWX (dual purpose embedding and infiltration parrafin) melting point 57c ? we tried switching, and it didn?t help.
- We only use distilled water in our baths
- Mercedes Starfrost charged slides
- Oven for 30 min at 62c
- We always use a cold red detection kit
-Sometimes our runs are delayed but not more than 4 hours
- Rundown is done in our stainer (standardized), and slides are coverslipped with tape.
- The bulk fluid containers in the bond are cleaned weekly and refrigerated between runs.
- The probe is cleaned every 300 slides.
- Mixing wells are cleaned out every two weeks and replaced often.
-Covertiles are well tended to.
We have tried to correct this problem, but we can't seem to fix it.
Any ideas?
https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_open-3Fid-3D1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA&d=DwICAg&c=yzGiX0CSJAqkDTmENO9LmP6KfPQitNABR9M66gsTb5w&r=ubybF7_hjNk08A-Q08Mf7WNmad5a9znkJq3XozlNVlg&m=UD2XLalsIAuK_lUZgavLUGnMMJaE8-g4Q28xJLvJLAw&s=kyqBaPGz-SvAoNDooiBtkSemt9dk4eOKo8l1XYvtmIs&e=<https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_open-3Fid-3D1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA&d=DwMFaQ&c=fHUKEfYp8ZKZp-Z4zyr9bXWWb-JCctOum5ZlzbjkVjM&r=1o7iZWfSCXEgDesmDMuV2rPUZJJnqv0nrvyFk2x0v0A&m=i7Ca5SrRojLnKG1c5k_oFTuocpFcJZ4BUGjdsiw477Q&s=v-vNoy8KgrPg0AxG3-Oxmp7s0DKlExpBrpdNhGkUWQ8&e=>
Jason R. Rosen, D.O.
Dermatopathologist
Premier Dermatology Partners
4675 Linton Boulevard, Suite 203
Delray Beach, FL 33445
(p) 561-499-5341
(f) 561-499-5343
(e) JRosen at totalderms.com<mailto:JRosen at totalderms.com>
------------------------------
Message: 2
Date: Tue, 9 Oct 2018 17:46:13 +0000 (UTC)
From: Rene J Buesa <rjbuesa at yahoo.com>
To: Linda Margraf <Linda.Margraf at cookchildrens.org>,
"histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] FW: Leica Bond Max artifact
Message-ID: <13494929.7046711.1539107173240 at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8
Dear Dr. Rosen:
I see nothing wrong in your protocol but your photo 0950 = 09501 seems as if there are 2 superimposed sections, one with a lot of brown particles? Is this artifact you are referring to? Seems "too organized" to be an artifact but it is brown and not red.
These "non DAB" reagents sometimes develop a sort of precipitate even within their expiration date. Why have you decided to use this red reagent instead of DAB? That could be a better option.
Other than that I am also completely baffled by your problem.
Ren?
On Tuesday, October 9, 2018 1:15 PM, Linda Margraf via Histonet <histonet at lists.utsouthwestern.edu> wrote:
Here is a message I am posting for Dr. Rosen:
Hello,
I am a longtime admirer of this list, and I am sure someone on here can help us out
We have an artifact on our immunohistochemistry (IHC) slides.
I will attempt to attach pictures using the upload tool. In case that doesn?t work, I am attaching a google drive link to some jpgs. The artifact is usually golden brown to red/pink in color, is clustered, appears both on top of the tissue and around it. If we do an aggressive rundown, it appears less red/brown and bluer.
We are using a Leica Bond MAX IHC system which is about a year old. We usually do two runs a day and 95% of the time we only use a red detection kit. We only see this on our red detection kit on our Leica bond. The brown is unaffected. The corresponding H&E slides are fine.
We started having this artifact after a month or two of production. 85% of the time we have some sort of artifact. Sometimes its perfect, sometimes it makes the tissue unreadable. We haven?t figured out any precipitating factors.
We have tried a number of things, but we still can?t get control of this.
Here is a brief overview of our protocols:
- Processor: Leica ASP300s tissue processor.
-Parrafin = Mercedes HWWX (dual purpose embedding and infiltration parrafin) melting point 57c ? we tried switching, and it didn?t help.
- We only use distilled water in our baths
- Mercedes Starfrost charged slides
- Oven for 30 min at 62c
- We always use a cold red detection kit
-Sometimes our runs are delayed but not more than 4 hours
- Rundown is done in our stainer (standardized), and slides are coverslipped with tape.
- The bulk fluid containers in the bond are cleaned weekly and refrigerated between runs.
- The probe is cleaned every 300 slides.
- Mixing wells are cleaned out every two weeks and replaced often.
-Covertiles are well tended to.
We have tried to correct this problem, but we can't seem to fix it.
Any ideas?
https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_open-3Fid-3D1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA&d=DwICAg&c=yzGiX0CSJAqkDTmENO9LmP6KfPQitNABR9M66gsTb5w&r=ubybF7_hjNk08A-Q08Mf7WNmad5a9znkJq3XozlNVlg&m=UD2XLalsIAuK_lUZgavLUGnMMJaE8-g4Q28xJLvJLAw&s=kyqBaPGz-SvAoNDooiBtkSemt9dk4eOKo8l1XYvtmIs&e=<https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_open-3Fid-3D1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA&d=DwMFaQ&c=fHUKEfYp8ZKZp-Z4zyr9bXWWb-JCctOum5ZlzbjkVjM&r=1o7iZWfSCXEgDesmDMuV2rPUZJJnqv0nrvyFk2x0v0A&m=i7Ca5SrRojLnKG1c5k_oFTuocpFcJZ4BUGjdsiw477Q&s=v-vNoy8KgrPg0AxG3-Oxmp7s0DKlExpBrpdNhGkUWQ8&e=>
Jason R. Rosen, D.O.
Dermatopathologist
Premier Dermatology Partners
4675 Linton Boulevard, Suite 203
Delray Beach, FL 33445
(p) 561-499-5341
(f) 561-499-5343
(e) JRosen at totalderms.com<mailto:JRosen at totalderms.com>
_______________________________________________
Histonet mailing list
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https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonet&d=DwICAg&c=yzGiX0CSJAqkDTmENO9LmP6KfPQitNABR9M66gsTb5w&r=ubybF7_hjNk08A-Q08Mf7WNmad5a9znkJq3XozlNVlg&m=UD2XLalsIAuK_lUZgavLUGnMMJaE8-g4Q28xJLvJLAw&s=65VTIbTt5Cez3DHVF4vIBCoqH0MsYay3VSIywTmpZfw&e=
------------------------------
Message: 3
Date: Tue, 9 Oct 2018 10:49:58 -0700
From: P Sicurello <patpxs at gmail.com>
To: HistoNet <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] H&E for H. pylori?
Message-ID:
<CAPSjddanVyjpNUvGfFWyNF86vhyDjbe8hFGSiSa6cLxuV4On9w at mail.gmail.com>
Content-Type: text/plain; charset="UTF-8"
Good Morning Listers,
Has anyone out there optimized an H&E for H. pylori?
Sure we can run a giemsa or and IHC but what fun is that?
Send me your ideas and make the day of our GI pathologists.
Thanks oodles.
Sincerely,
Paula Sicurello, HTL (ASCP)CM
Histotechnology Specialist
UC San Diego Health
9300 Campus Point Drive
La Jolla, CA 92037
(P): 858-249-5610
*Confidentiality Notice*: The information transmitted in this e-mail is
intended only for the person or entity to which it is addressed and may
contain confidential and/or privileged material. Any review,
retransmission, dissemination or other use of or taking of any action in
reliance upon this information by persons or entities other than the
intended recipient is prohibited. If you received this e-mail in error,
please contact the sender and delete the material from any computer.
------------------------------
Message: 4
Date: Tue, 9 Oct 2018 19:46:01 +0000 (UTC)
From: Rene J Buesa <rjbuesa at yahoo.com>
To: P Sicurello <patpxs at gmail.com>, HistoNet
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] H&E for H. pylori?
Message-ID: <1870025542.7160799.1539114361850 at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8
Paula:"Optimizing" H&E to detect H. piloris is a rout you should not attempt?because H&E cannot be "optimized" for that.Giemsa is?one way or, even better, modified Steiner stain. The only disadvantage modified Steiner has?is using radioactive uranium nitrate, but I developed a procedure where 0.02%?aq. phosphotungstic acid solution can be used instead.?IHC will be way more expensive.
Ren?
On Tuesday, October 9, 2018 2:03 PM, P Sicurello via Histonet <histonet at lists.utsouthwestern.edu> wrote:
Good Morning Listers,
Has anyone out there optimized an H&E for H. pylori?
Sure we can run a giemsa or and IHC but what fun is that?
Send me your ideas and make the day of our GI pathologists.
Thanks oodles.
Sincerely,
Paula Sicurello, HTL (ASCP)CM
Histotechnology Specialist
UC San Diego Health
9300 Campus Point Drive
La Jolla, CA 92037
(P): 858-249-5610
*Confidentiality Notice*: The information transmitted in this e-mail is
intended only for the person or entity to which it is addressed and may
contain confidential and/or privileged material.? Any review,
retransmission, dissemination or other use of or taking of any action in
reliance upon this information by persons or entities other than the
intended recipient is prohibited.? If you received this e-mail in error,
please contact the sender and delete the material from any computer.
_______________________________________________
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------------------------------
Message: 5
Date: Tue, 9 Oct 2018 22:59:15 +0000
From: "Tony Henwood (SCHN)" <tony.henwood at health.nsw.gov.au>
To: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] H&E for H. pylori?
Message-ID:
<b4cbab7946d242fa92170c32c74cc2ba at SVDCMBX-MEX024.nswhealth.net>
Content-Type: text/plain; charset="utf-8"
The following might be useful:
Tazawa, K., & Tsutsumi, Y. (1998). Effect of prolonged staining with hematoxylin on detecting Helicobacter pyiori in hematoxylin?eosin?stained gastric mucosa. Pathology international, 48(6), 448-452.
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: Rene J Buesa via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Wednesday, 10 October 2018 6:46 AM
To: P Sicurello; HistoNet
Subject: Re: [Histonet] H&E for H. pylori?
Paula:"Optimizing" H&E to detect H. piloris is a rout you should not attempt?because H&E cannot be "optimized" for that.Giemsa is?one way or, even better, modified Steiner stain. The only disadvantage modified Steiner has?is using radioactive uranium nitrate, but I developed a procedure where 0.02%?aq. phosphotungstic acid solution can be used instead.?IHC will be way more expensive.
Ren?
On Tuesday, October 9, 2018 2:03 PM, P Sicurello via Histonet <histonet at lists.utsouthwestern.edu> wrote:
Good Morning Listers,
Has anyone out there optimized an H&E for H. pylori?
Sure we can run a giemsa or and IHC but what fun is that?
Send me your ideas and make the day of our GI pathologists.
Thanks oodles.
Sincerely,
Paula Sicurello, HTL (ASCP)CM
Histotechnology Specialist
UC San Diego Health
9300 Campus Point Drive
La Jolla, CA 92037
(P): 858-249-5610
*Confidentiality Notice*: The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material.? Any review, retransmission, dissemination or other use of or taking of any action in reliance upon this information by persons or entities other than the intended recipient is prohibited.? If you received this e-mail in error, please contact the sender and delete the material from any computer.
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Message: 6
Date: Tue, 9 Oct 2018 19:33:06 -0400
From: Erin Sarricks <esarricks at gmail.com>
To: histonet <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Job Opening in Massachusetts: IHC Lab Supervisor
Message-ID:
<CAPapK_1kPNq6u3VWWHJQHZL0XGA1cg9hJQdMf7wqgy7tUAR17g at mail.gmail.com>
Content-Type: text/plain; charset="UTF-8"
HSRL (Histo-Scientific Research Labs) is a leading GLP contract research
organization which specializes in veterinary histopathology since 1999. Our
staff has been committed to scientific integrity and superior
responsiveness to client needs. A career with HSRL offers you potential to
join an established company with exceptional growth opportunity.
HSRL is seeking a qualified Immunohistochemistry Laboratory Supervisor to
join our team at our facility in Worcester, MA.
This position is split between bench histology/IHC and managing a small
team of specialists.
Essential Functions and Responsibilities:
- Perform routine histology, including tissue processing, fixation,
embedding, sectioning, staining, and imaging with the highest of quality.
- Oversee GLP IHC program.
- Develop Immunohistochemistry stain protocols and adhere to write SOPs.
- Effectively collaborate with team members and clients
- Various administrative duties to include recording and maintaining
detailed records
- Excellent organizational skills and ability to multi-task
- Works independently and troubleshoots technical problems
Qualifications:
- Minimum 3 years of related experience as a histologic technician with
concentration in veterinary histopathology and IHC techniques.
- ASCP certified Histotechnician (HT) or Histotechnologist (HTL) or
eligible, certification preferred.
- Demonstrated knowledge of and skill in oral and written communication,
problem solving, adaptability, and teamwork.
- Excellent organizational skills and ability to multi-task.
- Experience with Biocare?s IntelliPATH Autostainer is preferred.
A comprehensive benefits package will be offered to the successful
candidate, salary commensurate with experience.
To learn more about HSRL, please visit https://urldefense.proofpoint.com/v2/url?u=http-3A__www.hsrl.org&d=DwICAg&c=yzGiX0CSJAqkDTmENO9LmP6KfPQitNABR9M66gsTb5w&r=ubybF7_hjNk08A-Q08Mf7WNmad5a9znkJq3XozlNVlg&m=UD2XLalsIAuK_lUZgavLUGnMMJaE8-g4Q28xJLvJLAw&s=A01xuw7HuwFrJIsZpyy99R_tWx6s6vclF1gInPKKpsw&e=
Job Type: Full-time
Experience:
- employment as a Histo Tech w/concentration in IHC techniques: 3 years
(Required)
------------------------------
Message: 7
Date: Tue, 9 Oct 2018 23:34:13 +0000
From: "Cooper, Brian" <bcooper at chla.usc.edu>
To: "'histonet at lists.utsouthwestern.edu'"
<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Low Oxygen Sensor/Alarms
Message-ID: <f5bfc32b6f104a77bf3722e9143b1e40 at chla.usc.edu>
Content-Type: text/plain; charset=us-ascii
Hi,
Looking at the newly revised LABGEN checklist this afternoon. GEN.77550 states "In areas where liquid nitrogen is used, there are oxygen sensors with a low oxygen alarm mounted in an appropriate location and sufficient airflow to prevent asphyxiation." For those of you who already do this, can you please tell me the type and model of sensor you're using?
Thanks,
Brian D. Cooper, HT (ASCP)CM | Histology Supervisor
Department of Pathology and Laboratory Medicine
Children's Hospital Los Angeles
4650 Sunset Blvd MS#43- Los Angeles, CA 90027
bcooper at chla.usc.edu<mailto:bcooper at chla.usc.edu>
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