[Histonet] Re: Histonet Digest, Vol 125, Issue 20
DAVID OSIAGWU
dosiagwu <@t> unilag.edu.ng
Thu Apr 17 22:52:16 CDT 2014
Hi Gayle, i saw your response to Merissa, quite good. i have also been having problems with lectin IHC and will appreciate it if you can also forward me your Lectin IHC protocol, and your recipe for the Lectin buffer.
Thank you
Regards.
Daniel
Daniel Osiagwu, MS
Dept. of Anatomic and Molecular Pathology
College of Medicine.
University of Lagos.
+2348188225457
dosiagwu <@t> unilag.edu.ng
----- Original Message -----
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To: histonet <@t> lists.utsouthwestern.edu
Sent: Thursday, April 17, 2014 5:49:21 PM
Subject: Histonet Digest, Vol 125, Issue 20
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Today's Topics:
1. RE: Staining FFPE with biotinylated SNA - how to block?
(James Watson)
2. Tracking control blocks (Fawn Bomar)
3. RE: Staining FFPE with biotinylated SNA - how to block?
(gayle callis)
4. RE: Tracking control blocks (Cartun, Richard)
5. RE: Xylene testing ANP 08216 (Michael LaFriniere)
6. RE: IHC imaging systems (Michael LaFriniere)
7. RE: IHC imaging systems (joelle weaver)
8. RE: Waste Flammable Info Please (Michael LaFriniere)
9. human cells in rat tissue (Marti Gaudes, Merce)
10. RE: human cells in rat tissue (Elizabeth Chlipala)
11. Billing Question (Adesupo, Adesuyi (Banjo))
----------------------------------------------------------------------
Message: 1
Date: Wed, 16 Apr 2014 18:15:58 +0000
From: James Watson <JWatson <@t> gnf.org>
Subject: [Histonet] RE: Staining FFPE with biotinylated SNA - how to
block?
To: "koellingr <@t> comcast.net" <koellingr <@t> comcast.net>, M.O.
<modz9636 <@t> gmail.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <A032B25305EC6949AC9AFF23C9620616C9F04655 <@t> EX5.lj.gnf.org>
Content-Type: text/plain; charset="utf-8"
With our lectin staining on paraffin embedded tissue we use Avidin/Biotin blocking prior to antibody application. Some protocols we do use HIER before the blocking. One important item is that some lectins require a specific diluent. The dilution can be as high as 1:12,000. Lectins are well known for binding with multiple tissue entities, sometimes you can reduce the staining on unwanted tissue entities by reducing your incubation time or dilution. We do not use serum blocking.
For PNA we use:
PNA Diluent
Calcium Chloride���������������������..��������������������� 0.0111 gm.
Magnesium Chloride��������������������������������������� 0.0203 gm.
Manganese Chloride��������������������������������������� 0.0125 gm.
Distilled Water���������������������������������������������.���. 100.0 ml
James Watson HT�� ASCP
GNF�� Genomics Institute of the Novartis Research Foundation
Tel������ 858-332-4647
Fax���� 858-812-1915
jwatson <@t> gnf.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of koellingr <@t> comcast.net
Sent: Wednesday, April 16, 2014 9:53 AM
To: M.O.
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Staining FFPE with biotinylated SNA - how to block?
Hi Merissa,
don't know if you got any private idea responses so I'll throw in my opinion.�� I would always worry about some of the things you are mentioning and that are standard thoughts regarding biotin block, retrieval, etc in IHC.
But I would think about your serum, which I steadfastly avoided with SNA or any lectin I used.���� Lectins look at glyco components and serum (or serum substitutes) can be full of glycoproteins and the target then is the blocking serum for your lectin which can cause bad background.�� I did and would use washes, diluents, etc that had NO serum or milk or anything like that in them.�� You can make your own, completely free of potentially having��glycoproteins or Vector sells some.�� For some lectins (look at a list of target sugars) you maybe can get by with serum or milk and such to block��but many I've found you just can't.
Ray (still in, whoever would have guessed, once again rainy Seattle)
----- Original Message -----
From: "M.O." <modz9636 <@t> gmail.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, April 15, 2014 5:59:51 PM
Subject: [Histonet] Staining FFPE with biotinylated SNA - how to block?
Hello Histonet!
I ran a trial on FFPE mouse samples with a biotinylated lectin, SNA from vector. ��The SNA is Biotinylated Sambucus Nigra Lectin (Elderberry). ��I have never stained with anything like this, so I ran a test.
I deparaffinized, blocked with NGS, incubated overnight at 4C with the diluted biotinylated SNA. ��On the second day, I used Vector's ABC kit and alkaline phosphatase (red) kit.
Once stained, I noticed a lot of background. ��After looking into the blocking step, would a biotin/avidin blocking step be the correct step instead of a serum because I don't have a secondary? ��How do I know what needs to be blocked - biotin, avidin or both? ��Is there a way to do this without a kit and use solutions I may have in my lab?
Thank you for your help!
Sincerely,
Merissa
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------------------------------
Message: 2
Date: Wed, 16 Apr 2014 18:30:00 +0000
From: Fawn Bomar <Fawn.Bomar <@t> HalifaxRegional.com>
Subject: [Histonet] Tracking control blocks
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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<0111BC10D77DC54EAB99B2DDA3BCE4B96F5FA5 <@t> EXCH-2K10.hrhs.com>
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Happy Easter everyone!!! I was wondering if anyone had a tracking system that they would be willing to share in regards to how you all are keeping a record of your control blocks for IHC. As of right now, we are cutting a slide and staining it for the pathologist to approve- then keeping the paperwork and slide in files, and putting a label on the block in use with the date of approval.
Thank you
Fawn
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Message: 3
Date: Wed, 16 Apr 2014 15:13:57 -0600
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: RE: [Histonet] Staining FFPE with biotinylated SNA - how to
block?
To: "Histonet " <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <003801cf59b8$c9509630$5bf1c290$@bresnan.net>
Content-Type: text/plain; charset="UTF-8"
Dear Merissa,
Ray is correct about using serums with lectins. In fact you really are NOT doing immunostaining unless you are using an antibody made to recognize that lectin e.g. anti-lectin therefore a normal serum block is not needed. Serums in fact contain glycoproteins that bind to lectins, so BSA is a better protein carrier for buffers and diluents if you want to make up your own. We used Jacksons immunoglobulin protease free BSA. Ray gave good advice on blocking techniques. However, if you do anti-Lectin, you would be doing IHC and then blocking is legal. If you want a protocol for lectin IHC, I will be happy to forward via private email. With some lectins for direct staining, you should use the Lectin buffer that has no phosphate ions, as explained by Vector technical services. I also have a recipe for the Lectin buffer if you want it. Avidin/biotin blocking is still needed with biotinylated lectin, particularly is the tissue is known to have endogenous biotin.
Be sure to find out if your lectin is sensitive to phosphate ions, or use TBS or the Lectin buffer.
Also, if you are doing a non-IHC direct lectin-biotin staining, you must do the correct negative control. For SNA, this is an elution with 500 mM lactose in buffered saline followed by 500 mM lactose in acetic acid to finish elution. Buffer alone is NOT a negative control. For the lectins we worked with, we diluted the lectin (working concentration) in the recommended mM inhibition sugar and let it sit in the refrigerator overnight, warmed to RT just before use as negative control. This allows the lectin to bind to its specific sugar, and not to glycoproteins in the tissue, but keeps the biotin, and in our case, fluorophore in the negative control.
There is an excellent, inexpensive book, Lectin Histochemistry, a Concise Practical Handbook by SA Brooks, AJC Leathem and U Schumacher that tells all about using many lectins, protocols for lectin IHC and lectin direct binding staining. An interesting side history is the founder of Vector is a lectin expert.
For those doing IHC with anti-lectin, antigen retrieval may be needed per James Watson reply and is included in the Lectin IHC protocol I have. I am presuming he was doing true IHC for his lectin work.
Gayle Callis
HTL/HT/MT(ASCP)
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of koellingr <@t> comcast.net
Sent: Wednesday, April 16, 2014 10:53 AM
To: M.O.
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Staining FFPE with biotinylated SNA - how to block?
Hi Merissa,
don't know if you got any private idea responses so I'll throw in my opinion. I would always worry about some of the things you are mentioning and that are standard thoughts regarding biotin block, retrieval, etc in IHC.
But I would think about your serum, which I steadfastly avoided with SNA or any lectin I used. Lectins look at glyco components and serum (or serum substitutes) can be full of glycoproteins and the target then is the blocking serum for your lectin which can cause bad background. I did and would use washes, diluents, etc that had NO serum or milk or anything like that in them. You can make your own, completely free of potentially having glycoproteins or Vector sells some. For some lectins (look at a list of target sugars) you maybe can get by with serum or milk and such to block but many I've found you just can't.
Ray (still in, whoever would have guessed, once again rainy Seattle)
----- Original Message -----
From: "M.O." <modz9636 <@t> gmail.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, April 15, 2014 5:59:51 PM
Subject: [Histonet] Staining FFPE with biotinylated SNA - how to block?
Hello Histonet!
I ran a trial on FFPE mouse samples with a biotinylated lectin, SNA from vector. The SNA is Biotinylated Sambucus Nigra Lectin (Elderberry). I have never stained with anything like this, so I ran a test.
I deparaffinized, blocked with NGS, incubated overnight at 4C with the diluted biotinylated SNA. On the second day, I used Vector's ABC kit and alkaline phosphatase (red) kit.
Once stained, I noticed a lot of background. After looking into the blocking step, would a biotin/avidin blocking step be the correct step instead of a serum because I don't have a secondary? How do I know what needs to be blocked - biotin, avidin or both? Is there a way to do this without a kit and use solutions I may have in my lab?
Thank you for your help!
Sincerely,
Merissa
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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------------------------------
Message: 4
Date: Thu, 17 Apr 2014 01:03:21 +0000
From: "Cartun, Richard" <Richard.Cartun <@t> hhchealth.org>
Subject: [Histonet] RE: Tracking control blocks
To: Fawn Bomar <Fawn.Bomar <@t> HalifaxRegional.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<9215BD4B0BA1B44D962A71C758B68D2E022AB2C7 <@t> HHCEXCHMB03.hhcsystem.org>
Content-Type: text/plain; charset="us-ascii"
The vast majority of our positive control tissues are prepared from "left-over" tissue not used for patient diagnosis. I am hesitant to use the diagnostic tissue for control purposes in case we have to go back to perform additional testing. Also, I try to identify cases where we can put-through at least 20-30 tissue blocks. All control cases are entered into a "FileMaker Pro" file that I created years ago. Each case is given a "C" number and we enter the actual case number, the specimen site, the diagnosis, the time and date that the tissue is placed in formalin, the date that the tissue is processed, and immunoreactivity data (e.g., CK-MNF116+, HER2+, ALK1+, etc.). This program allows us to search any field using free text; very helpful when identifying a case for control purposes. All of our positive control slides have the "C" number on them.
Richard
Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
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.cartun <@t> hhchealth.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Fawn Bomar
Sent: Wednesday, April 16, 2014 2:30 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Tracking control blocks
Happy Easter everyone!!! I was wondering if anyone had a tracking system that they would be willing to share in regards to how you all are keeping a record of your control blocks for IHC. As of right now, we are cutting a slide and staining it for the pathologist to approve- then keeping the paperwork and slide in files, and putting a label on the block in use with the date of approval.
Thank you
Fawn
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Thank you
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Message: 5
Date: Thu, 17 Apr 2014 12:48:58 +0000
From: Michael LaFriniere <Michael.LaFriniere <@t> ccplab.com>
Subject: [Histonet] RE: Xylene testing ANP 08216
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4A2A16B9707CE04E9CB6C82DC18C1D29669F24 <@t> AHCMSASEXCH02.my.ahc.local>
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All of my labs are tested at least yearly or when we change a major procedure dealing with formalin or xylene. In addition if an associate feels the fumes are high we test that day. When I CAP inspect I like to see the lab tests annually, however, it's not the requirement but would recommend as a caution to demonstrating a safe environment.
Michael R. LaFriniere, HT (ASCP)
Executive Director
Capital Choice Pathology Laboratory
12041 Bournefield Way, Suite A * Silver Spring, MD 20904
P: 240.471.3427 * F: 240.471.3401 * Cell 410-940-8844
michael.lafriniere <@t> CCPLab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala
Sent: Tuesday, April 15, 2014 11:41 AM
To: Piche, Jessica; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Xylene testing ANP 08216
We test every year.
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 <@t> premierlab.com
www.premierlab.com
March 10, 2014 is Histotechnology Professionals Day
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Piche, Jessica
Sent: Tuesday, April 15, 2014 9:33 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Xylene testing ANP 08216
Hey Everyone,
Just curious what people are doing with xylene testing? Are people really only doing the initial testing and never testing again if all was good? Seems like there should be periodic testing after decades have gone by!
Thanks,
Jessica Piche, HT(ASCP)
Waterbury Hospital
CONFIDENTIALITY NOTICE: This email and any attachments contain confidential information that is legally privileged. This information is intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing this information to any other party unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of these documents is strictly prohibited. If you have received this information in error, please notify the sender immediately and delete these documents. Copyright (c) Waterbury Hospital _______________________________________________
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------------------------------
Message: 6
Date: Thu, 17 Apr 2014 12:57:37 +0000
From: Michael LaFriniere <Michael.LaFriniere <@t> ccplab.com>
Subject: RE: [Histonet] IHC imaging systems
To: Paula Sicurello <patpxs <@t> gmail.com>, HistoNet
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4A2A16B9707CE04E9CB6C82DC18C1D29669F5D <@t> AHCMSASEXCH02.my.ahc.local>
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I am setting up the Ventana system and performing validation, it's taking a little longer to set up then expected but the support has been excellent. Hopefully will have it running and validated in the next few months. Validation is the time consumer I found with setting up a system.
Michael R. LaFriniere, HT (ASCP)
Executive Director
Capital Choice Pathology Laboratory
12041 Bournefield Way, Suite A * Silver Spring, MD 20904
P: 240.471.3427 * F: 240.471.3401 * Cell 410-940-8844
michael.lafriniere <@t> CCPLab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Sicurello
Sent: Tuesday, April 15, 2014 12:59 PM
To: HistoNet
Subject: [Histonet] IHC imaging systems
Hello out there in Histoland,
If any of you out there in the clinical environment are using imaging
systems to quantify IHC slides, please let me know what system you are
using.
We are currently using a system that was provided through Dako, but they no
longer support it. So time to consider buying a new one. It needs to have
an algorithm that allows the system to distinguish between labelled and not
labelled cells and the percentage of each.
Thanks in advance,
Paula
--
Paula Sicurello, HTL (ASCP)
Supervisor, Clinical Electron Microscopy Laboratory
Duke University Health System
Rm.#251M, Duke South, Green Zone
Durham, North Carolina 27710
P: 919.684.2091
HIPAA Privacy Notification: This message and any accompanying documents are
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and contain information intended for the specific individual (s) only. This
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hereby notified that you have received this document in error and that any
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------------------------------
Message: 7
Date: Thu, 17 Apr 2014 13:02:36 +0000
From: joelle weaver <joelleweaver <@t> hotmail.com>
Subject: RE: [Histonet] IHC imaging systems
To: Michael LaFriniere <michael.lafriniere <@t> ccplab.com>, Paula
Sicurello <patpxs <@t> gmail.com>, HistoNet
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <SNT149-W15520234221F0CFBED82ACD8520 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
Amen... the time investment for validation!
Celebrate when it is completed :)
Joelle Weaver MAOM, HTL (ASCP) QIHC
> From: Michael.LaFriniere <@t> ccplab.com
> To: patpxs <@t> gmail.com; histonet <@t> lists.utsouthwestern.edu
> Date: Thu, 17 Apr 2014 12:57:37 +0000
> Subject: RE: [Histonet] IHC imaging systems
> CC:
>
> I am setting up the Ventana system and performing validation, it's taking a little longer to set up then expected but the support has been excellent. Hopefully will have it running and validated in the next few months. Validation is the time consumer I found with setting up a system.
>
> Michael R. LaFriniere, HT (ASCP)
> Executive Director
>
> Capital Choice Pathology Laboratory
> 12041 Bournefield Way, Suite A * Silver Spring, MD 20904
> P: 240.471.3427 * F: 240.471.3401 * Cell 410-940-8844
> michael.lafriniere <@t> CCPLab.com
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Paula Sicurello
> Sent: Tuesday, April 15, 2014 12:59 PM
> To: HistoNet
> Subject: [Histonet] IHC imaging systems
>
> Hello out there in Histoland,
>
> If any of you out there in the clinical environment are using imaging
> systems to quantify IHC slides, please let me know what system you are
> using.
>
> We are currently using a system that was provided through Dako, but they no
> longer support it. So time to consider buying a new one. It needs to have
> an algorithm that allows the system to distinguish between labelled and not
> labelled cells and the percentage of each.
>
> Thanks in advance,
>
> Paula
>
> --
> Paula Sicurello, HTL (ASCP)
> Supervisor, Clinical Electron Microscopy Laboratory
> Duke University Health System
> Rm.#251M, Duke South, Green Zone
> Durham, North Carolina 27710
> P: 919.684.2091
>
> HIPAA Privacy Notification: This message and any accompanying documents are
> covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521,
> and contain information intended for the specific individual (s) only. This
> information is confidential. If you are not the intended recipient or an
> agent responsible for delivering it to the intended recipient, you are
> hereby notified that you have received this document in error and that any
> review, dissemination, copying or the taking of any action based on the
> contents of this information is strictly prohibited . If you have received
> this communication in error, please notify us immediately by e-mail, and
> delete the original message.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
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------------------------------
Message: 8
Date: Thu, 17 Apr 2014 13:12:19 +0000
From: Michael LaFriniere <Michael.LaFriniere <@t> ccplab.com>
Subject: [Histonet] RE: Waste Flammable Info Please
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4A2A16B9707CE04E9CB6C82DC18C1D29669F96 <@t> AHCMSASEXCH02.my.ahc.local>
Content-Type: text/plain; charset="us-ascii"
I found each State that I have been in has a little different method to hazardous waste handling/reporting in the laboratory. I have found that recycling alcohol, xylene and formalin greatly reduces cost; my labs demonstrate 60-70% cost reduction in all areas of concern, (usage, disposal and regulation costs). However, you must invest time for a successful "program", that includes ample space, training and monitoring to assure its maximum benefit. In today's environment of decreasing laboratory reimbursement this is a major "LEAN" process that can be highly valuable in the pathology laboratory!
Michael R. LaFriniere, HT (ASCP)
Executive Director
Capital Choice Pathology Laboratory
12041 Bournefield Way, Suite A * Silver Spring, MD 20904
P: 240.471.3427 * F: 240.471.3401 * Cell 410-940-8844
michael.lafriniere <@t> CCPLab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala
Sent: Tuesday, April 15, 2014 1:21 PM
To: Dennis Hahn; 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] RE: Waste Flammable Info Please
Dennis
First of all we are in Colorado. If you dispose of more than 650 gallons a year in waste you are considered a small quantity generator, that amounts to one 55 gallon drum a month. Once you become a SQG this requires being registered with the EPA, additional safety training, every other year on site audits (In Colorado the Colorado Department of Public Health takes care of this) and then the other year a self-certification check list needs to be completed. If you dispose of less than 650 gallons you are considered a conditionally exempt small quantity generator and are not required to be registered with the EPA. Since you are disposing of 1 to 2 - 55 gallon drums a month that would mean that you would need to decrease your waste by at most 660 gallons a year or by 50%.
When we recycled only alcohol in 2013 it looked like we still had around 1 - 55 gallon of waste per month. In 2014 We started recycling alcohol, xylene and proper, so far this year we have only had 2 - 55 gallons of waste picked up, that accounts for a 50% decrease in total waste, so it might be possible for you to start recycling and decrease your waste stream by 50%. We have also seen a cost saving in the purchasing of both alcohol and xylene the amount we purchase has decreased I don't have the specifics on the amount I just know we order less often that we used to.
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 <@t> premierlab.com
www.premierlab.com
March 10, 2014 is Histotechnology Professionals Day
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Dennis Hahn
Sent: Tuesday, April 15, 2014 10:55 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Waste Flammable Info Please
I have an inquiry specifically for Texas and/or Children's hospitals:
What is your current volume of waste flammables and how do you handle it? Are you recycling, or is it removed by an outside vendor? A combination of the two? Suggestions on recycling instrumentation? Any drawbacks?
We currently collect all waste flammable materials into a 55 Gallon drum and it is removed once or twice a month by a contracted vendor. The problem is, we are disposing of so much now that the vendor is requiring the medical center to obtain a high-use license involving the state and the EPA. We have evaluated recycling many years ago and found the process to be slow and sort of cumbersome. I know that the process has improved quite a bit over the years, but what about the tech time, tech exposure, time for recycling, etc.? Have you found it to be highly cost effective?
Thanks in advance for any info you can share.
Dennis
Dennis Hahn, HT (ASCP)
Histology Lab Supervisor
Cook Children's Medical Center
801 7th Avenue
Ft. Worth, TX 76104
(682) 885-6133
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Message: 9
Date: Thu, 17 Apr 2014 14:41:08 +0000
From: "Marti Gaudes, Merce" <mmarti <@t> cmrb.eu>
Subject: [Histonet] human cells in rat tissue
To: "Histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <3d47706b1b9f4e1898dc4d7bc68b799f <@t> CMRBMAIL.cmrb.name>
Content-Type: text/plain; charset="iso-8859-1"
Hi, good afternoon everyone,
I was wondering if anybody could help me out. We would like to detect human cells engrafted in rat tissue, particularly in formalin-fixed, paraffin-embedded tissue. I have so far tried one anti-human nuclear antigen antibody that works well in criostat...but without good results in paraffin.
Do you know any anti human nuclear antibody that works well in paraffin? Or...do you know any other protein that let me differentiate between human cells and rat tissue? Obviously my cells are not GFP positive...
Thanks a lot!
Merc� Mart� Gaudes
Histology Unit
________________________________
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Message: 10
Date: Thu, 17 Apr 2014 08:53:06 -0600
From: Elizabeth Chlipala <liz <@t> premierlab.com>
Subject: [Histonet] RE: human cells in rat tissue
To: "Marti Gaudes, Merce" <mmarti <@t> cmrb.eu>,
"Histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<14E2C6176416974295479C64A11CB9AE019C79E05916 <@t> SBS2K8.premierlab.local>
Content-Type: text/plain; charset="iso-8859-1"
Merce
There is another Human Nuclear antibody from cell signaling we find that it works in FFPE tissues, however in our hands it did not stain all nuclei probably around 90- 95% overall. We found it to be potentially fixation dependent, we noticed that tissues that had spent longer time in fixative did not stain that well and we also saw some staining in some rat samples, we could not figure out if it was due to our protocol or tissue collection but we decided that there was too many inconsistencies so we did not use it for that particular study. We might try it again. We do use another antibody to detect human cells in a rat background and that is HLA-1 the one we use is from abcam.
Good Luck
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
(303) 682-9060 fax
(303) 881-0763 cell
liz <@t> premierlab.com
www.premierlab.com
March 10, 2014 is Histotechnology Professionals Day
Ship to Address:
Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Marti Gaudes, Merce
Sent: Thursday, April 17, 2014 8:41 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] human cells in rat tissue
Hi, good afternoon everyone,
I was wondering if anybody could help me out. We would like to detect human cells engrafted in rat tissue, particularly in formalin-fixed, paraffin-embedded tissue. I have so far tried one anti-human nuclear antigen antibody that works well in criostat...but without good results in paraffin.
Do you know any anti human nuclear antibody that works well in paraffin? Or...do you know any other protein that let me differentiate between human cells and rat tissue? Obviously my cells are not GFP positive...
Thanks a lot!
Merc� Mart� Gaudes
Histology Unit
________________________________
P Abans d'imprimir aquest missatge, si us plau, comprova que �s realment necessari. El medi ambient �s cosa de tots.
La informaci� continguda en aquest missatge i en qualsevol fitxer adjunt �s confidencial, privada i d'�s exclusiu per al destinatari. Si no �s la persona a la qual anava dirigida aquesta informaci�, si us plau, notifiqui immediatament l'enviament erroni al remitent i esborri el missatge. Qualsevol c�pia, divulgaci�, distribuci� o utilitzaci� noautoritzada d'aquest correu electr�nic i dels seus adjunts est� prohibida en virtut de la legislaci� vigent.
P Antes de imprimir este mensaje, por favor, comprueba que es realmente necesario. El medio ambiente es cosa de todos.
La informaci�n contenida en este mensaje y en cualquier fichero adjunto es confidencial, privada y de uso exclusivo para el destinatario. Si usted no es la persona a la cual iba dirigida esta informaci�n, por favor, notifique inmediatamente el envio err�neo al remitente y borre el mensaje. Cualquier copia, divulgaci�n, distribuci�n o utilizaci�n no autorizada de este correo electr�nico y de sus adjuntos est� prohibida en virtud de la legislaci�n vigente.
P Before printing this e-mail, please make certain it is absolutely necessary. The environment is everybody's business.
The information included in this e-mail and any attached files is confidential and private. If you are not the intended recipient, please notify the sender and delete this message immediately. Dissemination, forwarding or copying of this e-mail and its associated attachments is strictly prohibited in accordance with current legislation.
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------------------------------
Message: 11
Date: Thu, 17 Apr 2014 11:19:44 -0500
From: "Adesupo, Adesuyi (Banjo)" <abadesuyi <@t> nrh-ok.com>
Subject: [Histonet] Billing Question
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<04EE4F75BB5FB246ADB68D69B74604438E3B8220E3 <@t> MAIL.nrhnt.nrh-ok.com>
Content-Type: text/plain; charset="us-ascii"
Hi,
Please I have a question on the new IHC Billing Policies for CPT, Medicare. My question is how do we go about the billing of the HER-2 DUAL ISH and the Kappa and Lambda ISH?
Thanks,
Banjo Adesuyi, BSMT, HT (ASCP) HTL, QIHC, QLS
Histology Supervisor
Norman Regional Health System,
Norman, OK 73071.
Tel: 405- 307- 1145
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End of Histonet Digest, Vol 125, Issue 20
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