Re: [Histonet] Hematoxylin Precipitate

[Manfre, Philip via Histonet]

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@lists.utsouthwestern.edu] 
Sent: Tuesday, September 22, 2015 5:04 PM
To: gayle.cal...@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.cal...@bresnan.net
> To: thiggin...@msn.com; histonet@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 

[Histonet] Cryostate decon ANP.23410

[Nancy Schmitt via Histonet]

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





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Re: [Histonet] Cryostate decon ANP.23410

[Michael Ann Jones via Histonet]

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
mjo...@metropath.com






On 9/23/15, 9:33 AM, "Nancy Schmitt via Histonet"
 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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Re: [Histonet] Hematoxylin Precipitate

[Bob Richmond via Histonet]

The alum hematoxylins I've worked with required daily filtration. An
iridescent scum on top of the hematoxylin means you're overdue to filter.

When you brew a fresh batch of alum hematoxylin, it's a light yellow color.
As it oxidizes (slowly if you depend on atmospheric oxygen, instantly if
you add an oxidant such sodium iodate or mercuric oxide) it turns into deep
purple hematein (say hee-muh-TEE-in, not the same as hematin) which is what
you stain with. Hematein oxidizes further to filterable crud.

Whatman No.1 filter paper is fast and easy to get, and is usually
preferred. Sometimes Good Management Practice requires substituting a paper
towel, which works reasonably well.

You young whippersnappers shouldn't need to be told this by a 76 year old
pathologist!

Bob Richmond
Samurai Pathologist
Maryville TN
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Re: [Histonet] RUO expiration

[Kiran via Histonet]

Dear Experts, 
How do you guys determine expiration dates for RUO Abs? Thx 
Kiran 

> On Sep 23, 2015, at 10:53 AM, Chung, Fai via Histonet 
>  wrote:
> 
> 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-
> From: histonet-requ...@lists.utsouthwestern.edu 
> [mailto:histonet-requ...@lists.utsouthwestern.edu]
> Sent: Wednesday, September 23, 2015 10:00 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: Histonet Digest, Vol 142, Issue 19
> 
> 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
> 
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>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: 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 Gillformulations

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Re: [Histonet] Ventana Discovery

[Chung, Fai via Histonet]

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-
From: histonet-requ...@lists.utsouthwestern.edu 
[mailto:histonet-requ...@lists.utsouthwestern.edu]
Sent: Wednesday, September 23, 2015 10:00 AM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 142, Issue 19

Send Histonet mailing list submissions to
histonet@lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
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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: 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 Gillformulations
  (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 
To: "histonet@lists.utsouthwestern.edu"

Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID: 
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" 
> To: "Histonet (histonet@lists.utsouthwestern.edu)"
>   
> Subject: [Histonet] Hematoxylin Precipitate
> Message-ID: <4cda87133587e64c965ce6c356d18...@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 
To: Tim H , "'histonet@lists.utsouthwestern.edu'
(histonet@lists.utsouthwestern.edu)"

Subject: Re: [Histonet] Hematoxylin Precipitate
Message-ID:
<14E2C6176416974295479C64A11CB9AE02230F9D9510@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 

[Histonet] Kappa/Lambda bone marrow

[Burnett, Brandy via Histonet]

Does anyone out there have a Kappa and Lambda protocol for ISH on the Ventana 
BenchMark Ultra?
Was also wondering what type of decal should be used for ISH?

Thanks!

Brandy Burnett
BS,HTL(ASCP) QIHC
bburn...@capecodhealth.org


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[Histonet] Fungus contamination on AFB slides

[Scott, Allison D via Histonet]

Hello to all in histoland.   We are currently having a problem with candida on 
our AFB slides.  We are doing the stain by hand due to the unavailability of 
the AFB kits from ventana.  We are using bottled sterile water for rinses and 
the slides are not being put in coplin jars for staining.  All solutions are 
being pipetted on to the slides.  All alcohols are fresh and the kit which is 
from American MasterTech is new, just opened.  This by the way is a cytology 
case(cystospin). Containers to run down to water have been bleached.  Any 
suggetions where to go next.


Allison Scott
LBJ Hospital
Houston, TExas
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