[Histonet] Ventana Discovery
Chung, Fai
Fai.Chung at cshs.org
Wed Sep 23 12:53:48 CDT 2015
Ventana is selling their Discovery instrument for research use only. I am wondering if anyone know if a complete full validation is done on the instrument, can we use it on clinical cases as a "laboratory developed test". For antibody classified as RUO, we are able to use it on patient slide as a "laboratory developed" test as long as we do a complete full validation.
Thanks.
Fai
-----Original Message-----
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Sent: Wednesday, September 23, 2015 10:00 AM
To: histonet at lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 142, Issue 19
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Today's Topics:
1. Re: Hematoxylin Precipitate (Tim H)
2. Re: Hematoxylin Precipitate (Elizabeth Chlipala)
3. Re: Hematoxylin Precipitate (Manfre, Philip)
4. Re: Hematoxylin Precipitate (Gayle Callis)
5. Webinar presentation hand-outs (Elaine allison Hoffman)
6. Re: Webinar presentation hand-outs (Morken, Timothy)
7. Re: Hematoxylin Precipitate (Tim H)
8. Elastic Stain on the Benchmark Special Stainer (BMSS)?
(P Sicurello)
9. Re: Hematoxylin Precipitate (Elizabeth Chlipala)
10. Quick H202 quenching question. (Lewis, Patrick)
11. Re: Hematoxylin Precipitate and filtering Gill formulations
(Gayle Callis)
12. Re: Hematoxylin Precipitate (Manfre, Philip)
13. Cryostate decon ANP.23410 (Nancy Schmitt)
14. Re: Cryostate decon ANP.23410 (Michael Ann Jones)
----------------------------------------------------------------------
Message: 1
Date: Tue, 22 Sep 2015 12:25:22 -0500
From: Tim H <thigginsht at msn.com>
To: "histonet at lists.utsouthwestern.edu"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID: <BAY179-W927F0A1854FFE1CA2310D0D8450 at phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
You should be filtering your Hematoxylin on a daily basis regardless of what the manufactures says. We use to filter twice a day since we did a traditional overnight run and then again in the afternoon for specimens that had been microwave processed. So much tissue washes off in the solutions they should be changed or filtered fairly regularly to try and prevent cross contamination on the slides.
You can also try increasing your rinse times and see if that doesn't help as well.
Thanks,
Tim
>
> Message: 1
> Date: Mon, 21 Sep 2015 15:14:39 -0500
> From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> To: "Histonet (histonet at lists.utsouthwestern.edu)"
> <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] Hematoxylin Precipitate
> Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello all,
>
> Has anyone using Richard Allen Hematoxylin-2 noticed an odd artifact on the slides after using the Hematoxylin for more than a few days on their stainer? We are seeing small spore or pollen-like blue dots here and there on the slides. It is not coming from the water bath or our water supply on the stainer. I used sterile gloves, opened a new case of slides, dipped them in DI water, then in the RA Hematoxylin 2 on the stainer, then in DI again, air-dried and coverslipped them, and the blue dots were there. The only way we got rid of the blue artifact was to use new RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
>
> Thanks for your input, and if you can recommend a different, reasonably priced hematoxylin, that would be awesome.
>
> Cheers,
>
> Sandy
>
>
>
> Sandra J. Cheasty, HT (ASCP)
>
> Histology & Necropsy Supervisor
>
> UW-Madison, School of Veterinary Medicine
------------------------------
Message: 2
Date: Tue, 22 Sep 2015 11:55:05 -0600
From: Elizabeth Chlipala <liz at premierlab.com>
To: Tim H <thigginsht at msn.com>, "'histonet at lists.utsouthwestern.edu'
(histonet at lists.utsouthwestern.edu)"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID:
<14E2C6176416974295479C64A11CB9AE02230F9D9510 at SBS2K8.premierlab.local>
Content-Type: text/plain; charset="us-ascii"
I think you may a have a serious problem if "so much tissue washes off" if that is happening you have problems with tissue adherence. A properly processed and cut section should not wash off the slide, or even a portion of it should not wash off. That would mean that you did not provide to the pathologist what is represented in the block. If that is the case then you would need to filter all solutions on a stainer daily not just the hematoxylin.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz at premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 11:25 AM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Hematoxylin Precipitate
You should be filtering your Hematoxylin on a daily basis regardless of what the manufactures says. We use to filter twice a day since we did a traditional overnight run and then again in the afternoon for specimens that had been microwave processed. So much tissue washes off in the solutions they should be changed or filtered fairly regularly to try and prevent cross contamination on the slides.
You can also try increasing your rinse times and see if that doesn't help as well.
Thanks,
Tim
>
> Message: 1
> Date: Mon, 21 Sep 2015 15:14:39 -0500
> From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> To: "Histonet (histonet at lists.utsouthwestern.edu)"
> <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] Hematoxylin Precipitate
> Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello all,
>
> Has anyone using Richard Allen Hematoxylin-2 noticed an odd artifact on the slides after using the Hematoxylin for more than a few days on their stainer? We are seeing small spore or pollen-like blue dots here and there on the slides. It is not coming from the water bath or our water supply on the stainer. I used sterile gloves, opened a new case of slides, dipped them in DI water, then in the RA Hematoxylin 2 on the stainer, then in DI again, air-dried and coverslipped them, and the blue dots were there. The only way we got rid of the blue artifact was to use new RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
>
> Thanks for your input, and if you can recommend a different, reasonably priced hematoxylin, that would be awesome.
>
> Cheers,
>
> Sandy
>
>
>
> Sandra J. Cheasty, HT (ASCP)
>
> Histology & Necropsy Supervisor
>
> UW-Madison, School of Veterinary Medicine
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Tue, 22 Sep 2015 14:26:04 -0400
From: "Manfre, Philip" <philip_manfre at merck.com>
To: Elizabeth Chlipala <liz at premierlab.com>, Tim H
<thigginsht at msn.com>
Cc: "'histonet at lists.utsouthwestern.edu'
(histonet at lists.utsouthwestern.edu)"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID:
<558A4571351D0C42BD923F403F4198C40102EF9B24B4 at USCTMXP51014.merck.com>
Content-Type: text/plain; charset="us-ascii"
Wow, I agree with Liz. There should not routinely be "so much tissue washing off". There is a fundamental problem, if this is the case.
With regards to hematoxylin, have you tried Gill's Hematoxylin, 1, 2, or 3? These do not need filtering and do not produce a precipitate.
Phil.
Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486
215-652-9750
215-993-0383 (fax)
philip_manfre at merck.com
-----Original Message-----
From: Elizabeth Chlipala via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 1:55 PM
To: Tim H; 'histonet at lists.utsouthwestern.edu' (histonet at lists.utsouthwestern.edu)
Subject: Re: [Histonet] Hematoxylin Precipitate
I think you may a have a serious problem if "so much tissue washes off" if that is happening you have problems with tissue adherence. A properly processed and cut section should not wash off the slide, or even a portion of it should not wash off. That would mean that you did not provide to the pathologist what is represented in the block. If that is the case then you would need to filter all solutions on a stainer daily not just the hematoxylin.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz at premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 11:25 AM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Hematoxylin Precipitate
You should be filtering your Hematoxylin on a daily basis regardless of what the manufactures says. We use to filter twice a day since we did a traditional overnight run and then again in the afternoon for specimens that had been microwave processed. So much tissue washes off in the solutions they should be changed or filtered fairly regularly to try and prevent cross contamination on the slides.
You can also try increasing your rinse times and see if that doesn't help as well.
Thanks,
Tim
>
> Message: 1
> Date: Mon, 21 Sep 2015 15:14:39 -0500
> From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> To: "Histonet (histonet at lists.utsouthwestern.edu)"
> <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] Hematoxylin Precipitate
> Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello all,
>
> Has anyone using Richard Allen Hematoxylin-2 noticed an odd artifact on the slides after using the Hematoxylin for more than a few days on their stainer? We are seeing small spore or pollen-like blue dots here and there on the slides. It is not coming from the water bath or our water supply on the stainer. I used sterile gloves, opened a new case of slides, dipped them in DI water, then in the RA Hematoxylin 2 on the stainer, then in DI again, air-dried and coverslipped them, and the blue dots were there. The only way we got rid of the blue artifact was to use new RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
>
> Thanks for your input, and if you can recommend a different, reasonably priced hematoxylin, that would be awesome.
>
> Cheers,
>
> Sandy
>
>
>
> Sandra J. Cheasty, HT (ASCP)
>
> Histology & Necropsy Supervisor
>
> UW-Madison, School of Veterinary Medicine
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
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Message: 4
Date: Tue, 22 Sep 2015 12:27:21 -0600
From: "Gayle Callis" <gayle.callis at bresnan.net>
To: "'Tim H'" <thigginsht at msn.com>, "Histonet"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID: <000601d0f564$523ae2f0$f6b0a8d0$@bresnan.net>
Content-Type: text/plain; charset="us-ascii"
Sandy,
After years of using Richard Allan's hematoxylin 2 with great success, if
we didn't filter daily before use, we had stain precipitate on sections.
Some of this comes from the hematoxylin continuing to oxidize in open air,
bacteria and other "crud". Tim is absolutely correct ignoring
manufacturers no filtering instructions. Being old school, we were taught
to faithfully filter any hematoxylin, regardless of progressive or
regressive types. If we topped off hematoxylin 2 or used new stock, the
stain was filtered into a CLEAN staining container/dish. Keep an extra
container around if possible. We used a medium fast filter paper, Whatman
54. I realize this takes time but junk on a slide is NOT good thing,
especially after IHC staining and have a photo to show this - the result of
being lazy and not filtering the hematoxylin on that particular day.
We used a distilled water rinse before hematoxylin2, but DI H2O will be
contaminated with cellular debris and last hydrating alcohol carryover.
Change DI water frequently if you have many runs in a day. We used 1
minute running tap water rinses after hematoxylin, clearant and bluing. If
you are using running water rinses, take a look at the blue ppt in the post
hematoxylin container as you don't want that sticking to sections. Non
running water rinses should be changed after each H&E run in my opinion.
Adequate clean water rinses are important to not have carry over of clearant
into bluing reagents or bluing reagent into eosin in order to maintain
correct pH for staining.
Good luck
Gayle M. Callis
HTL/HT/MT(ASCP)
-----Original Message-----
From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 11:25 AM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Hematoxylin Precipitate
You should be filtering your Hematoxylin on a daily basis regardless of what
the manufactures says. We use to filter twice a day since we did a
traditional overnight run and then again in the afternoon for specimens that
had been microwave processed. So much tissue washes off in the solutions
they should be changed or filtered fairly regularly to try and prevent cross
contamination on the slides.
You can also try increasing your rinse times and see if that doesn't help as
well.
Thanks,
Tim
>
> Message: 1
> Date: Mon, 21 Sep 2015 15:14:39 -0500
> From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> To: "Histonet (histonet at lists.utsouthwestern.edu)"
> <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] Hematoxylin Precipitate
> Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello all,
>
> Has anyone using Richard Allen Hematoxylin-2 noticed an
odd artifact on the slides after using the Hematoxylin for more than a few
days on their stainer? We are seeing small spore or pollen-like blue dots
here and there on the slides. It is not coming from the water bath or our
water supply on the stainer. I used sterile gloves, opened a new case of
slides, dipped them in DI water, then in the RA Hematoxylin 2 on the
stainer, then in DI again, air-dried and coverslipped them, and the blue
dots were there. The only way we got rid of the blue artifact was to use new
RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
>
> Thanks for your input, and if you can recommend a
different, reasonably priced hematoxylin, that would be awesome.
>
> Cheers,
>
> Sandy
>
>
>
> Sandra J. Cheasty, HT (ASCP)
>
> Histology & Necropsy Supervisor
>
> UW-Madison, School of Veterinary Medicine
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Tue, 22 Sep 2015 18:44:33 +0000 (UTC)
From: Elaine allison Hoffman <elaineahoffman55 at yahoo.com>
To: Histonet List <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Webinar presentation hand-outs
Message-ID:
<1367610594.1779042.1442947473127.JavaMail.yahoo at mail.yahoo.com>
Content-Type: text/plain; charset=UTF-8
Hi all,
Just recently joined in with an Advance webinar last week, 9/15/15, Tuesday with speaker, Tim Morken on "Basic Principles of Validation in Histology".? The webinar was very informative and thorough.? The power-point presentation looked to be about 40 pages long.? I'm interested in getting a copy of the the presentation shown during the webinar but don't know how.? There was definitely too much to remember and our lab could really use it ASAP. Wasn't able to write it down fast enough.? I left emails with Advance but no answer back yet.? Any suggestions??? Appreciate any help, thanks....
Elaine Hoffman, HT(ASCP)
The Gastroenterology Clinic & Endoscopy Center, Inc.GI Pathology Associates
------------------------------
Message: 6
Date: Tue, 22 Sep 2015 19:17:43 +0000
From: "Morken, Timothy" <Timothy.Morken at ucsf.edu>
To: Elaine allison Hoffman <elaineahoffman55 at yahoo.com>
Cc: Histonet <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Webinar presentation hand-outs
Message-ID:
<761E2B5697F795489C8710BCC72141FF603161D9 at ex07.net.ucsf.edu>
Content-Type: text/plain; charset="utf-8"
Elaine you can download the powerpoint and all the handouts from the Advance for Laboratories website under Webinars, QA/QC, " Basic Principles of Validation in Histology"
Link is below:
http://laboratory-manager.advanceweb.com/Webinar/Editorial-Webinars/Basic-Principles-of-Validation-in-Histology.aspx
I hope you find it helpful. Contact me directly with any questions or comments.
Tim Morken
Pathology Site Manager, Parnassus
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center
-----Original Message-----
From: Elaine allison Hoffman via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 11:45 AM
To: Histonet List
Subject: [Histonet] Webinar presentation hand-outs
Hi all,
Just recently joined in with an Advance webinar last week, 9/15/15, Tuesday with speaker, Tim Morken on "Basic Principles of Validation in Histology".? The webinar was very informative and thorough.? The power-point presentation looked to be about 40 pages long.? I'm interested in getting a copy of the the presentation shown during the webinar but don't know how.? There was definitely too much to remember and our lab could really use it ASAP. Wasn't able to write it down fast enough.? I left emails with Advance but no answer back yet.? Any suggestions??? Appreciate any help, thanks....
Elaine Hoffman, HT(ASCP)
The Gastroenterology Clinic & Endoscopy Center, Inc.GI Pathology Associates
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 7
Date: Tue, 22 Sep 2015 16:04:18 -0500
From: Tim H <thigginsht at msn.com>
To: "gayle.callis at bresnan.net" <gayle.callis at bresnan.net>, Histonet
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID: <BAY179-W211F093E50F381732F5A2ED8450 at phx.gbl>
Content-Type: text/plain; charset="Windows-1252"
Liz, Phillip and all that are
interested,
I take it you have guys never looked at or had someone else
examine what is at the bottom of the Hematoxylin filter after
you put through a day?s work. There will be tissue particles of
tissue along with other contaminates, I am not saying you are going to see
a complete LEEP sitting at the bottom but you will have
contamination.
Liz, maybe the tissue super adheres up there in wacky tabacky country
and Phillip sitting in a research facility (are you even
involved with processing and staining of slides or just publishing
articles?) but in the rest of the United States, I can guarantee you there
will be some tissue in the Hematoxylin if you use a traditional dip and
dunk system.
Liz, you do bring up a good point
about having tissue in every container. To a degree you
will have tissue in every reagent container. I was assuming and maybe
unjustly that most labs are using Good Laboratory Practice and
discard their reagents after they have used them for a staining session. This
topic was about Hematoxylin Precipitate and "small spore or pollen-like
blue dots" which I did not say was tissue, I was passing along some
of my 23 years of knowledge.
Listen; don't take my word for
it. You can have Ventana come out to your facility and filter
all your reagents and stains and have them tell you what is in your
solutions.
Next time just pass along good
information and criticize those trying to help! To many people like to make a negative on this site of people trying to truly help. This is why I rarely post on Histonet but instead directly email the person.
By no means am I promoting
Ventana/Roche products. I am just passing along information that might be
helpful.
Tim Disclaimer: This information is by no means is meant for any weak stomach individuals or those preparing for the zombie apocalypse.
> From: gayle.callis at bresnan.net
> To: thigginsht at msn.com; histonet at lists.utsouthwestern.edu
> Subject: RE: [Histonet] Hematoxylin Precipitate
> Date: Tue, 22 Sep 2015 12:27:21 -0600
>
> Sandy,
>
> After years of using Richard Allan's hematoxylin 2 with great success, if
> we didn't filter daily before use, we had stain precipitate on sections.
> Some of this comes from the hematoxylin continuing to oxidize in open air,
> bacteria and other "crud". Tim is absolutely correct ignoring
> manufacturers no filtering instructions. Being old school, we were taught
> to faithfully filter any hematoxylin, regardless of progressive or
> regressive types. If we topped off hematoxylin 2 or used new stock, the
> stain was filtered into a CLEAN staining container/dish. Keep an extra
> container around if possible. We used a medium fast filter paper, Whatman
> 54. I realize this takes time but junk on a slide is NOT good thing,
> especially after IHC staining and have a photo to show this - the result of
> being lazy and not filtering the hematoxylin on that particular day.
>
> We used a distilled water rinse before hematoxylin2, but DI H2O will be
> contaminated with cellular debris and last hydrating alcohol carryover.
> Change DI water frequently if you have many runs in a day. We used 1
> minute running tap water rinses after hematoxylin, clearant and bluing. If
> you are using running water rinses, take a look at the blue ppt in the post
> hematoxylin container as you don't want that sticking to sections. Non
> running water rinses should be changed after each H&E run in my opinion.
> Adequate clean water rinses are important to not have carry over of clearant
> into bluing reagents or bluing reagent into eosin in order to maintain
> correct pH for staining.
>
> Good luck
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
>
> -----Original Message-----
> From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
> Sent: Tuesday, September 22, 2015 11:25 AM
> To: histonet at lists.utsouthwestern.edu
> Subject: Re: [Histonet] Hematoxylin Precipitate
>
> You should be filtering your Hematoxylin on a daily basis regardless of what
> the manufactures says. We use to filter twice a day since we did a
> traditional overnight run and then again in the afternoon for specimens that
> had been microwave processed. So much tissue washes off in the solutions
> they should be changed or filtered fairly regularly to try and prevent cross
> contamination on the slides.
>
> You can also try increasing your rinse times and see if that doesn't help as
> well.
>
> Thanks,
>
> Tim
> >
> > Message: 1
> > Date: Mon, 21 Sep 2015 15:14:39 -0500
> > From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> > To: "Histonet (histonet at lists.utsouthwestern.edu)"
> > <histonet at lists.utsouthwestern.edu>
> > Subject: [Histonet] Hematoxylin Precipitate
> > Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Hello all,
> >
> > Has anyone using Richard Allen Hematoxylin-2 noticed an
> odd artifact on the slides after using the Hematoxylin for more than a few
> days on their stainer? We are seeing small spore or pollen-like blue dots
> here and there on the slides. It is not coming from the water bath or our
> water supply on the stainer. I used sterile gloves, opened a new case of
> slides, dipped them in DI water, then in the RA Hematoxylin 2 on the
> stainer, then in DI again, air-dried and coverslipped them, and the blue
> dots were there. The only way we got rid of the blue artifact was to use new
> RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
> >
> > Thanks for your input, and if you can recommend a
> different, reasonably priced hematoxylin, that would be awesome.
> >
> > Cheers,
> >
> > Sandy
> >
> >
> >
> > Sandra J. Cheasty, HT (ASCP)
> >
> > Histology & Necropsy Supervisor
> >
> > UW-Madison, School of Veterinary Medicine
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 8
Date: Tue, 22 Sep 2015 14:46:11 -0700
From: P Sicurello <patpxs at gmail.com>
To: HistoNet <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Elastic Stain on the Benchmark Special Stainer
(BMSS)?
Message-ID:
<CAPSjdda-VoNHtRK20upt8bowUZq_QCE66RwqDMwXwQF2v7fMJQ at mail.gmail.com>
Content-Type: text/plain; charset=UTF-8
Hi Netters,
Has anyone out in Histoland achieved blackness with the BMSS Elastic stain?
All I'm getting are purple elastic fibers. If you have crossed over to the
darker side of staining and have a protocol you'd be willing to share, by
all means let me know.
I want to join the darkside too! At least with the Elastic stain.
Thanks in advance,
Paula :-)
UCSD Health System
------------------------------
Message: 9
Date: Tue, 22 Sep 2015 16:21:17 -0600
From: Elizabeth Chlipala <liz at premierlab.com>
To: Tim H <thigginsht at msn.com>
Cc: "'histonet at lists.utsouthwestern.edu'
(histonet at lists.utsouthwestern.edu)"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID:
<14E2C6176416974295479C64A11CB9AE02230F9D951C at SBS2K8.premierlab.local>
Content-Type: text/plain; charset="us-ascii"
Tim
First of all my comment was not meant to criticize your post and even if you may not think so I was trying to help. I stand by what I said, my comment was to address what I thought was the amount of tissue loss your lab experiences and if I took your comment too literally I apologize but I firmly believe that properly processed and sectioned tissue samples should remain on the slide. I do understand that we will on occasion have loss of tissue from the slides that we cut and stain. We will see a lens floating in one of the alcohols when we stain mouse eyes or a portion of a dermal construct will come off the slides if we have not dried it properly. Most of the tissue loss we experience is due to improperly processed and sectioned samples.
We are a small lab and we are GLP compliant. We do not change our H&E staining reagents daily our volume varies depending upon the projects we are working on, but I can tell you that we have looked at H&E staining over time and we have made sure that our reagents are changed appropriately. We do not filter our hematoxylin daily and have not experienced many floaters or carry over or other things on our slides.
Liz from wacky tabacky country
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz at premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 3:04 PM
To: gayle.callis at bresnan.net; Histonet
Subject: Re: [Histonet] Hematoxylin Precipitate
Liz, Phillip and all that are
interested,
I take it you have guys never looked at or had someone else examine what is at the bottom of the Hematoxylin filter after you put through a day's work. There will be tissue particles of tissue along with other contaminates, I am not saying you are going to see a complete LEEP sitting at the bottom but you will have contamination.
Liz, maybe the tissue super adheres up there in wacky tabacky country and Phillip sitting in a research facility (are you even involved with processing and staining of slides or just publishing
articles?) but in the rest of the United States, I can guarantee you there will be some tissue in the Hematoxylin if you use a traditional dip and dunk system.
Liz, you do bring up a good point
about having tissue in every container. To a degree you will have tissue in every reagent container. I was assuming and maybe unjustly that most labs are using Good Laboratory Practice and discard their reagents after they have used them for a staining session. This topic was about Hematoxylin Precipitate and "small spore or pollen-like blue dots" which I did not say was tissue, I was passing along some of my 23 years of knowledge.
Listen; don't take my word for
it. You can have Ventana come out to your facility and filter all your reagents and stains and have them tell you what is in your solutions.
Next time just pass along good
information and criticize those trying to help! To many people like to make a negative on this site of people trying to truly help. This is why I rarely post on Histonet but instead directly email the person.
By no means am I promoting
Ventana/Roche products. I am just passing along information that might be helpful.
Tim Disclaimer: This information is by no means is meant for any weak stomach individuals or those preparing for the zombie apocalypse.
> From: gayle.callis at bresnan.net
> To: thigginsht at msn.com; histonet at lists.utsouthwestern.edu
> Subject: RE: [Histonet] Hematoxylin Precipitate
> Date: Tue, 22 Sep 2015 12:27:21 -0600
>
> Sandy,
>
> After years of using Richard Allan's hematoxylin 2 with great success, if
> we didn't filter daily before use, we had stain precipitate on sections.
> Some of this comes from the hematoxylin continuing to oxidize in open air,
> bacteria and other "crud". Tim is absolutely correct ignoring
> manufacturers no filtering instructions. Being old school, we were taught
> to faithfully filter any hematoxylin, regardless of progressive or
> regressive types. If we topped off hematoxylin 2 or used new stock, the
> stain was filtered into a CLEAN staining container/dish. Keep an extra
> container around if possible. We used a medium fast filter paper, Whatman
> 54. I realize this takes time but junk on a slide is NOT good thing,
> especially after IHC staining and have a photo to show this - the result of
> being lazy and not filtering the hematoxylin on that particular day.
>
> We used a distilled water rinse before hematoxylin2, but DI H2O will
> be contaminated with cellular debris and last hydrating alcohol carryover.
> Change DI water frequently if you have many runs in a day. We used 1
> minute running tap water rinses after hematoxylin, clearant and bluing. If
> you are using running water rinses, take a look at the blue ppt in the post
> hematoxylin container as you don't want that sticking to sections. Non
> running water rinses should be changed after each H&E run in my opinion.
> Adequate clean water rinses are important to not have carry over of
> clearant into bluing reagents or bluing reagent into eosin in order to maintain
> correct pH for staining.
>
> Good luck
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
>
> -----Original Message-----
> From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
> Sent: Tuesday, September 22, 2015 11:25 AM
> To: histonet at lists.utsouthwestern.edu
> Subject: Re: [Histonet] Hematoxylin Precipitate
>
> You should be filtering your Hematoxylin on a daily basis regardless
> of what the manufactures says. We use to filter twice a day since we
> did a traditional overnight run and then again in the afternoon for
> specimens that had been microwave processed. So much tissue washes
> off in the solutions they should be changed or filtered fairly
> regularly to try and prevent cross contamination on the slides.
>
> You can also try increasing your rinse times and see if that doesn't
> help as well.
>
> Thanks,
>
> Tim
> >
> > Message: 1
> > Date: Mon, 21 Sep 2015 15:14:39 -0500
> > From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> > To: "Histonet (histonet at lists.utsouthwestern.edu)"
> > <histonet at lists.utsouthwestern.edu>
> > Subject: [Histonet] Hematoxylin Precipitate
> > Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Hello all,
> >
> > Has anyone using Richard Allen Hematoxylin-2 noticed
> > an
> odd artifact on the slides after using the Hematoxylin for more than a
> few days on their stainer? We are seeing small spore or pollen-like
> blue dots here and there on the slides. It is not coming from the
> water bath or our water supply on the stainer. I used sterile gloves,
> opened a new case of slides, dipped them in DI water, then in the RA
> Hematoxylin 2 on the stainer, then in DI again, air-dried and
> coverslipped them, and the blue dots were there. The only way we got
> rid of the blue artifact was to use new RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
> >
> > Thanks for your input, and if you can recommend a
> different, reasonably priced hematoxylin, that would be awesome.
> >
> > Cheers,
> >
> > Sandy
> >
> >
> >
> > Sandra J. Cheasty, HT (ASCP)
> >
> > Histology & Necropsy Supervisor
> >
> > UW-Madison, School of Veterinary Medicine
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
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------------------------------
Message: 10
Date: Tue, 22 Sep 2015 22:36:09 +0000
From: "Lewis, Patrick" <patrick.lewis at seattlechildrens.org>
To: " (Histonet at lists.utsouthwestern.edu)"
<Histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Quick H202 quenching question.
Message-ID:
<3903BE18914F4440834F0E620415FFCA3CBA2228 at PPWEXD01d.childrens.sea.kids>
Content-Type: text/plain; charset="us-ascii"
If I have an IHC where I am staining 2 slides from the same block, one with one primary antibody and the other with a different primary antibody.
And one antibody's slides have high nonspecific background, but the other antibody's slides have no background, Can I deduce that the background staining is not caused by insufficient H202 quenching/blocking?
All the steps were the same, except for the primary antibody/secondary antibody. Also, they did have different epitope retrievals.
But the wash buffers/blocking buffers/and substrate were the same.
I am fairly confident that my background problems are related to this particular primary antibody, and not the actual quenching/blocknig from my IHC protocol, but I thought I'd check with you guys.
Patrick.
Patrick Lewis
Research Associate II Bench
Seattle Childrens Research Institute
206-884-1115
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------------------------------
Message: 11
Date: Tue, 22 Sep 2015 16:59:55 -0600
From: "Gayle Callis" <gayle.callis at bresnan.net>
To: "'Manfre, Philip'" <philip_manfre at merck.com>
Cc: "Histonet" <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate and filtering Gill
formulations
Message-ID: <000301d0f58a$663f49a0$32bddce0$@bresnan.net>
Content-Type: text/plain; charset="us-ascii"
Yes, I have used Gill 1, 2 and 3 even in the early days of buying these
formulations from a vendor, and always filtered them before using.
Old school habits never changed..............
Gayle Callis
-----Original Message-----
From: Manfre, Philip via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 12:26 PM
To: Elizabeth Chlipala <liz at premierlab.com>; Tim H <thigginsht at msn.com>
Cc: 'histonet at lists.utsouthwestern.edu' (histonet at lists.utsouthwestern.edu)
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Wow, I agree with Liz. There should not routinely be "so much tissue
washing off". There is a fundamental problem, if this is the case.
With regards to hematoxylin, have you tried Gill's Hematoxylin, 1, 2, or 3?
These do not need filtering and do not produce a precipitate.
Phil.
Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486
215-652-9750
215-993-0383 (fax)
philip_manfre at merck.com
------------------------------
Message: 12
Date: Wed, 23 Sep 2015 07:55:59 -0400
From: "Manfre, Philip" <philip_manfre at merck.com>
To: Tim H <thigginsht at msn.com>, "gayle.callis at bresnan.net"
<gayle.callis at bresnan.net>, "'histonet at lists.utsouthwestern.edu'
(histonet at lists.utsouthwestern.edu)"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID:
<558A4571351D0C42BD923F403F4198C40102EF9B26BF at USCTMXP51014.merck.com>
Content-Type: text/plain; charset="us-ascii"
Well Tim,
In the 31 years I have been in histology, I obviously have done all lab work with my eyes closed, never looking at the equipment or solutions I have been using. I guess my stint in a high throughput contract lab taught me nothing nor did my 25 + years in big Pharma, notorious for hiring every hack who knows what a microscope slide is. You suggested that I sit around getting published... I am published, BTW, but that accounts for 0.01% of my experiences in the lab, yes, in the lab processing, embedding, sectioning (quite good at that), cryosectioning, performing and troubleshooting IHC and histochemical stains.
Something I may have learned, and maybe you will when you catch up, is not assume you know what someone else's work experiences may have been or are. Histology occurs in many environments with differing equipment, reagents, processing styles, etc. Some are better than others. I guess our inadequate processing methods fall short, leaving far too much tissue on the slide where it belongs, and thus we don't have big tissue globs clogging everything, nor do we see precipitate with the hematoxylin used. Traditional formulations of hematoxylin, sure, absolutely filter. For anyone who benefits from filtering, why not continue. No harm will come. I was merely trying to advise someone who seemed to have an issue I thought I could help with. Now I realize all issues should be relegated to Tim, the Charles Churukian of our times. Look him up, if you're stumped by the reference. Maybe I'll head for Wacky Tabacky land for a vacation. It is quite beautiful there...
Phil.
-----Original Message-----
From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, September 22, 2015 5:04 PM
To: gayle.callis at bresnan.net; Histonet
Subject: Re: [Histonet] Hematoxylin Precipitate
Liz, Phillip and all that are
interested,
I take it you have guys never looked at or had someone else
examine what is at the bottom of the Hematoxylin filter after
you put through a day's work. There will be tissue particles of
tissue along with other contaminates, I am not saying you are going to see
a complete LEEP sitting at the bottom but you will have
contamination.
Liz, maybe the tissue super adheres up there in wacky tabacky country
and Phillip sitting in a research facility (are you even
involved with processing and staining of slides or just publishing
articles?) but in the rest of the United States, I can guarantee you there
will be some tissue in the Hematoxylin if you use a traditional dip and
dunk system.
Liz, you do bring up a good point
about having tissue in every container. To a degree you
will have tissue in every reagent container. I was assuming and maybe
unjustly that most labs are using Good Laboratory Practice and
discard their reagents after they have used them for a staining session. This
topic was about Hematoxylin Precipitate and "small spore or pollen-like
blue dots" which I did not say was tissue, I was passing along some
of my 23 years of knowledge.
Listen; don't take my word for
it. You can have Ventana come out to your facility and filter
all your reagents and stains and have them tell you what is in your
solutions.
Next time just pass along good
information and criticize those trying to help! To many people like to make a negative on this site of people trying to truly help. This is why I rarely post on Histonet but instead directly email the person.
By no means am I promoting
Ventana/Roche products. I am just passing along information that might be
helpful.
Tim Disclaimer: This information is by no means is meant for any weak stomach individuals or those preparing for the zombie apocalypse.
> From: gayle.callis at bresnan.net
> To: thigginsht at msn.com; histonet at lists.utsouthwestern.edu
> Subject: RE: [Histonet] Hematoxylin Precipitate
> Date: Tue, 22 Sep 2015 12:27:21 -0600
>
> Sandy,
>
> After years of using Richard Allan's hematoxylin 2 with great success, if
> we didn't filter daily before use, we had stain precipitate on sections.
> Some of this comes from the hematoxylin continuing to oxidize in open air,
> bacteria and other "crud". Tim is absolutely correct ignoring
> manufacturers no filtering instructions. Being old school, we were taught
> to faithfully filter any hematoxylin, regardless of progressive or
> regressive types. If we topped off hematoxylin 2 or used new stock, the
> stain was filtered into a CLEAN staining container/dish. Keep an extra
> container around if possible. We used a medium fast filter paper, Whatman
> 54. I realize this takes time but junk on a slide is NOT good thing,
> especially after IHC staining and have a photo to show this - the result of
> being lazy and not filtering the hematoxylin on that particular day.
>
> We used a distilled water rinse before hematoxylin2, but DI H2O will be
> contaminated with cellular debris and last hydrating alcohol carryover.
> Change DI water frequently if you have many runs in a day. We used 1
> minute running tap water rinses after hematoxylin, clearant and bluing. If
> you are using running water rinses, take a look at the blue ppt in the post
> hematoxylin container as you don't want that sticking to sections. Non
> running water rinses should be changed after each H&E run in my opinion.
> Adequate clean water rinses are important to not have carry over of clearant
> into bluing reagents or bluing reagent into eosin in order to maintain
> correct pH for staining.
>
> Good luck
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
>
> -----Original Message-----
> From: Tim H via Histonet [mailto:histonet at lists.utsouthwestern.edu]
> Sent: Tuesday, September 22, 2015 11:25 AM
> To: histonet at lists.utsouthwestern.edu
> Subject: Re: [Histonet] Hematoxylin Precipitate
>
> You should be filtering your Hematoxylin on a daily basis regardless of what
> the manufactures says. We use to filter twice a day since we did a
> traditional overnight run and then again in the afternoon for specimens that
> had been microwave processed. So much tissue washes off in the solutions
> they should be changed or filtered fairly regularly to try and prevent cross
> contamination on the slides.
>
> You can also try increasing your rinse times and see if that doesn't help as
> well.
>
> Thanks,
>
> Tim
> >
> > Message: 1
> > Date: Mon, 21 Sep 2015 15:14:39 -0500
> > From: "Sandra Cheasty" <cheastys at svm.vetmed.wisc.edu>
> > To: "Histonet (histonet at lists.utsouthwestern.edu)"
> > <histonet at lists.utsouthwestern.edu>
> > Subject: [Histonet] Hematoxylin Precipitate
> > Message-ID: <4cda87133587e64c965ce6c356d18f59 at svm.vetmed.wisc.edu>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Hello all,
> >
> > Has anyone using Richard Allen Hematoxylin-2 noticed an
> odd artifact on the slides after using the Hematoxylin for more than a few
> days on their stainer? We are seeing small spore or pollen-like blue dots
> here and there on the slides. It is not coming from the water bath or our
> water supply on the stainer. I used sterile gloves, opened a new case of
> slides, dipped them in DI water, then in the RA Hematoxylin 2 on the
> stainer, then in DI again, air-dried and coverslipped them, and the blue
> dots were there. The only way we got rid of the blue artifact was to use new
> RA Hematoxylin-2 every 2-3 days, which is a bit expensive.
> >
> > Thanks for your input, and if you can recommend a
> different, reasonably priced hematoxylin, that would be awesome.
> >
> > Cheers,
> >
> > Sandy
> >
> >
> >
> > Sandra J. Cheasty, HT (ASCP)
> >
> > Histology & Necropsy Supervisor
> >
> > UW-Madison, School of Veterinary Medicine
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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Message: 13
Date: Wed, 23 Sep 2015 15:33:30 +0000
From: Nancy Schmitt <Nancy_Schmitt at pa-ucl.com>
To: "'histonet at lists.utsouthwestern.edu'"
<histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Cryostate decon ANP.23410
Message-ID:
<906B4DA90ED1DB4DB6C7E94D7CEE6C360115A4B354 at PEITHA.wad.pa-ucl.com>
Content-Type: text/plain; charset="us-ascii"
Good Morning-
Defrost and decontaminate with TB disinfectant weekly if used daily. How are you best managing this and what are you using to decontaminate for TB?
Thank you
Nancy Schmitt MLT, HT(ASCP)
United Clinical Laboratories
Dubuque, IA 52001
NOTICE: This email may contain legally privileged information. The information
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------------------------------
Message: 14
Date: Wed, 23 Sep 2015 16:04:46 +0000
From: Michael Ann Jones <mjones at metropath.com>
To: Nancy Schmitt <Nancy_Schmitt at pa-ucl.com>,
"'histonet at lists.utsouthwestern.edu'"
<histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] Cryostate decon ANP.23410
Message-ID: <D22829A0.C8DC%mjones at metropath.com>
Content-Type: text/plain; charset="us-ascii"
We do maybe 11 frozens/month and so decon/defrost quarterly.
Michael Ann
Michael Ann Jones, HT (ASCP)
Histology Manager
Metropath
7444 W. Alaska Dr. #250
Lakewood, CO 80226
303.634.2511
Mjones at metropath.com
On 9/23/15, 9:33 AM, "Nancy Schmitt via Histonet"
<histonet at lists.utsouthwestern.edu> wrote:
>Good Morning-
>Defrost and decontaminate with TB disinfectant weekly if used daily. How
>are you best managing this and what are you using to decontaminate for TB?
>Thank you
>
>Nancy Schmitt MLT, HT(ASCP)
>United Clinical Laboratories
>Dubuque, IA 52001
>
>
>
>
>
>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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