[Histonet] RE:xylene substitute and marking pens
Price, Tiffany
Tiffany.Price <@t> thomaswv.org
Tue Jan 30 09:53:01 CST 2007
We use the Formula 83 substitute and really like it. It also recycles very well. We use Richard Allen mounting medium to coverslip- it is the most compatible, and recommended by CBG Biotech, the manufacturer of the clearant. We also use SHUR/MARK pens from Triangle Biomedical Sciences- they have a refillable tip that you just replace until the ink runs out of the pen. They come in red or black. I have found that these are the only pens that work with the Formula 83 clearant.
Hope this helps!
Tiffany
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Tuesday, January 30, 2007 10:05 AM
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Subject: Histonet Digest, Vol 38, Issue 46
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Today's Topics:
1. RE: Xylene substitute (sheila adey)
2. Re: Xylene substitute (Lynette Pavelich)
3. RE: pens (Bonner, Janet)
4. Miller's Elastic Stain (Andrea Grantham)
5. Hypoxia (Gamal Akabani)
6. RE: Miller's Elastic Stain (Monfils, Paul)
7. If you are even remotely curious, please call! Even more temp
openings-- (Cheryl Kerry)
8. Re: Miller's Elastic Stain (Bryan Hewlett)
9. Re: Miller's Elastic Stain (Bryan Llewellyn)
10. Re: Miller's Elastic Stain (Andrea Grantham)
11. Re: Anybody doing flow cytometry? What does Neat mean?
(Christina Thurby)
12. ASCP Exam (Stasha McCollum)
13. Processing debate (GMartin <@t> marshallhospital.org)
14. a question (nizar limaiem)
15. Re: ASCP Exam (Kaye Ryan)
16. human total IgG (Liz Chlipala)
17. RE: Re: Anybody doing flow cytometry? What does Neat mean?
(Monfils, Paul)
18. MCP1 positive control (Brown, Graham W)
19. Re: Mouse Testes Fixation and Processing (Laurie Reilly)
20. Re: Immunogold question (Robert Chiovetti)
21. Follow up (GMartin <@t> marshallhospital.org)
22. RE: Re: Anybody doing flow cytometry? What does Neat mean?
(Tarango, Mark)
23. ASCP EXAM (MICHELLE SEAGLE)
24. Re: Hypoxia (Jakub Otahal)
25. RUO's and diagnostic IHC (Houston, Ronald)
26. Re: Processing debate (Rene J Buesa)
----------------------------------------------------------------------
Message: 1
Date: Mon, 29 Jan 2007 13:38:44 -0500
From: "sheila adey" <sheila_adey <@t> hotmail.com>
Subject: RE: [Histonet] Xylene substitute
To: GMartin <@t> marshallhospital.org, histonet <@t> lists.utsouthwestern.edu
Message-ID: <BAY129-F110682FB4EE8B7B40FEFE393A70 <@t> phx.gbl>
Content-Type: text/plain; format=flowed
We tried using formula 88 and found it to be also irritating, especially
after a spill. It was a problem when we tried to coverslip because our
sugipath mounting media is Xylene based but since you have already covered
that issue, it would be nice to hear what you think of the quality of the
product. Please share after you've been using it.
Sheila Adey HT MLT
Port Huron Hospital
Michigan
>From: GMartin <@t> marshallhospital.org
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] Xylene substitute
>Date: Mon, 29 Jan 2007 09:15:00 -0800
>
>We presently use Xylene in our processor. We will be moving to a Xylene
>substitute. We are a small lab and still hand cover all of our slide. In
>the cover slip line we do use Sugipath's Sub-x Xylene substitute and are
>planning on using Sub-x.
>Can anyone in the group fill me in on their experience with a change over
>to a Xylene substitute.
>Thank you
>Gary Martin
>El Dorado pathology
>California.
>
>
>
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>Histonet <@t> lists.utsouthwestern.edu
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------------------------------
Message: 2
Date: Mon, 29 Jan 2007 13:41:25 -0500
From: "Lynette Pavelich" <lpaveli1 <@t> hurleymc.com>
Subject: Re: [Histonet] Xylene substitute
To: <histonet <@t> lists.utsouthwestern.edu>,<GMartin <@t> marshallhospital.org>
Message-ID: <45BDF985020000EE00010CF6 <@t> smtp-gw.hurleymc.com>
Content-Type: text/plain; charset=US-ASCII
Hi Gary,
We recenty had a change over to a substitute and went with the aliphatic
Propar by Anatech. Easy transition, very low oder, docs didn't notice
(big plus). We did add an extra container on the stainer when
depariffinizing, and on the processor, we have 3 containers of the
substitute. We are also able to recycle it. Works well with hand
coverslipping media also. You'll be happy with the change.
Lynette
Lynette Pavelich, HT(ASCP)
Histology Supervisor
Hurley Medical Center
One Hurley Plaza
Flint, MI 48503
ph: 810-257-9948
fax: 810-762-7082
>>> <GMartin <@t> marshallhospital.org> 01/29/07 12:15 PM >>>
We presently use Xylene in our processor. We will be moving to a Xylene
substitute. We are a small lab and still hand cover all of our slide.
In
the cover slip line we do use Sugipath's Sub-x Xylene substitute and are
planning on using Sub-x.
Can anyone in the group fill me in on their experience with a change
over
to a Xylene substitute.
Thank you
Gary Martin
El Dorado pathology
California.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Mon, 29 Jan 2007 14:51:14 -0500
From: "Bonner, Janet" <Janet.Bonner <@t> FLHOSP.ORG>
Subject: RE: [Histonet] pens
To: "Patricia Karlisch" <pkarlisch <@t> psu.edu>,
Histonet <@t> lists.utsouthwestern.edu, "ruegg, patsy"
<pruegg <@t> ihctech.net>, "Webb', 'Dorothy L"
<Dorothy.L.Webb <@t> HealthPartners.Com>
Message-ID:
<5F31F38C96781A4FBE3196EBC22D478004A73C <@t> fhosxchmb006.ADVENTISTCORP.NET>
Content-Type: text/plain; charset=iso-8859-1
We have been looking for a similar pen that will stay on the cassettes AND the slides. Right now we use two different pens. Smudging has been a problem, but the Mercedes pens seem like a winner!
Janet
________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Patricia Karlisch
Sent: Mon 1/29/2007 9:18 AM
To: Histonet <@t> lists.utsouthwestern.edu; ruegg, patsy; Webb', 'Dorothy L
Subject: RE: [Histonet] pens
All,
I have received good recommendations for cassette pens and it looks like there are many more pens out there then I knew about. We will try each one and see if this alleviates the smudging. Thank you all so much. Pat Karlisch.
Pat Karlisch
Supervisor, Histology, Pathology and Laboratory Medicine
Penn State Milton S. Hershey Medical Center
Mail Code H179
Hershey, PA 17033
Phone (717) 531-6072
Fax: (717) 531- 7741
email: pkarlisch <@t> psu.edu
*****E-Mail Confidentiality Notice*****
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>>> "patsy ruegg" <pruegg <@t> ihctech.net> 1/27/2007 7:44 PM >>>
We use pens from StatLabs we like, just have to let the cassettes dry a
minute or two before putting them in alcohol waiting for processing.
Patsy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patricia
Karlisch
Sent: Wednesday, January 24, 2007 8:54 AM
To: Dorothy L Webb; Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] pens
Dorothy,
Thank you for this information. We are looking for non-smudge pens
as well. This will help. Did you find that the Surgipath pens did not
smudge or come off during processing?
Pat Karlisch
Supervisor, Histology, Pathology and Laboratory Medicine
Penn State Milton S. Hershey Medical Center
Mail Code H179
Hershey, PA 17033
Phone (717) 531-6072
Fax: (717) 531- 7741
email: pkarlisch <@t> psu.edu
*****E-Mail Confidentiality Notice*****
This message (including any attachments) contains information intended
for a specific individual(s) and purpose that may be privileged,
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applicable law. Any inappropriate use, distribution or copying of the
message is strictly prohibited and may subject you to criminal or civil
penalty. If you have received this transmission in error, please reply
to the sender indicating this error and delete the transmission from
your system immediately.
>>> "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com> 1/24/2007
10:49 AM >>>
We recently had the same problems and researched several companies and
brands and found for our usage the following were the best:
Surgipath pens #01880 come in a box of 10 for $27.00
Marketlab pens #ML0479 come in a box of 10 for $37.00
Mercedes Medical has a new pen from Norway that we tried that are only
$13 per box of 10, but do not have the order number for them as they
are
not in stock quite yet.
The pens from Newcomer (Histo pen) and the Cancer Diagnostics pen did
not smear, etc., but dried out so fast and often came already dry in
the
box.
Good Luck!!
________________________________________
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------------------------------
Message: 4
Date: Mon, 29 Jan 2007 13:33:12 -0700
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Subject: [Histonet] Miller's Elastic Stain
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<4.3.2.7.2.20070129130923.00e1fec0 <@t> algranth.inbox.email.arizona.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Happy Monday!
I have had a request to do a Miller's Elastic Stain. I have almost all of
the stains and I do have all of the chemicals to do the stain but I have a
question.
One of the ingredients in the elastic stain is resorcin. I'm not a chemist
so I have to ask this question, I have resorcinol in my chemical cabinet -
is this the same thing as resorcin? I can find resorcinol in the Sigma cat.
but not resorcin.
Thanks!
Andi Grantham
.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist University of Arizona :
: (office: AHSC 4212) P.O. Box 245044 :
: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
------------------------------
Message: 5
Date: Mon, 29 Jan 2007 14:40:39 -0600
From: Gamal Akabani <gamal.akabani <@t> gmail.com>
Subject: [Histonet] Hypoxia
To: "Histonet ((E-mail))" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4885123C-E70A-41BC-AAE0-1F483B088D7A <@t> gmail.com>
Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed
Dear friends,
I am want to process some histological samples from glioma patients
in order to observe the level or grade of hypoxia on them, if any.
I am not a guru. Any help is appreciated on the methods or a
specific antibody used to visualize it. Thanks in advanced.
Gamal
------------------------------
Message: 6
Date: Mon, 29 Jan 2007 15:50:40 -0500
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] Miller's Elastic Stain
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4EBFF65383B74D49995298C4976D1D5E273C0E <@t> LSRIEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain; charset="iso-8859-1"
Yes, resorcinol and resorcin are the same thing, C6H6O2
------------------------------
Message: 7
Date: Mon, 29 Jan 2007 14:53:02 -0600
From: "Cheryl Kerry" <tkngflght <@t> yahoo.com>
Subject: [Histonet] If you are even remotely curious, please call!
Even more temp openings--
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <00b501c743e7$77b140c0$6401a8c0 <@t> FSDESKTOP>
Content-Type: text/plain; charset="US-ASCII"
Hi All!
We're swamped and need more great techs!! If you have ever been ever
remotely curious about travel temps, I would love to talk with you. Even if
you won't be ready for a year or more--let's start the conversation so when
you are ready to go, you're comfortable and happy in your decision.
I keep in touch. I don't pressure you (it is YOUR life, right?) Someone said
I don't come across as a used car salesman.I'm a histotech!! If something
doesn't fit--you tell me 'no' and we move on to the next opening. Easy as
that.
I have EIGHT open positions and need to fill them within the next two weeks.
All shifts, all skills. With the changes in the registry and state
licensing, we're going to continue to need great techs who want to try new
things.
Give me a call--it's an investment of about 15 minutes for the basics. I
help new travelers get started and we've got a great team. It all starts
with a question- "So, tell me about temping?"
I'll give you the details of the jobs that fit when we get a chance to
chat!!
Cheryl :-)
Cheryl Kerry, HT(ASCP)
Full Staff Inc
281.852.9457
800.756.3309
------------------------------
Message: 8
Date: Mon, 29 Jan 2007 15:54:58 -0500
From: "Bryan Hewlett" <bhewlett <@t> cogeco.ca>
Subject: Re: [Histonet] Miller's Elastic Stain
To: <histonet <@t> lists.utsouthwestern.edu>, "Andrea Grantham"
<algranth <@t> u.arizona.edu>
Message-ID: <000d01c743e7$bd2f0470$6500a8c0 <@t> mainbox>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=response
Andi,
Resorcin is the same thing as resorcinol!
Bryan
----- Original Message -----
From: "Andrea Grantham" <algranth <@t> u.arizona.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, January 29, 2007 3:33 PM
Subject: [Histonet] Miller's Elastic Stain
> Happy Monday!
> I have had a request to do a Miller's Elastic Stain. I have almost all of
> the stains and I do have all of the chemicals to do the stain but I have a
> question.
> One of the ingredients in the elastic stain is resorcin. I'm not a chemist
> so I have to ask this question, I have resorcinol in my chemical cabinet -
> is this the same thing as resorcin? I can find resorcinol in the Sigma
> cat. but not resorcin.
>
> Thanks!
> Andi Grantham
> .....................................................................
> : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
> : Sr. Research Specialist University of Arizona :
> : (office: AHSC 4212) P.O. Box 245044 :
> : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
> : (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
> :...................................................................:
> http://www.cba.arizona.edu/histology-lab.html
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 9
Date: Mon, 29 Jan 2007 12:43:26 -0800
From: Bryan Llewellyn <llewllew <@t> shaw.ca>
Subject: Re: [Histonet] Miller's Elastic Stain
To: Histonet <histonet <@t> lists.utsouthwestern.edu>, Andrea Grantham
<algranth <@t> u.arizona.edu>
Message-ID: <001201c743e6$209f0520$6400a8c0 <@t> yourlk4rlmsu>
Content-Type: text/plain; format=flowed; charset=iso-8859-1;
reply-type=response
Same thing.
Bryan Llewellyn
----- Original Message -----
From: "Andrea Grantham" <algranth <@t> u.arizona.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, January 29, 2007 12:33 PM
Subject: [Histonet] Miller's Elastic Stain
> Happy Monday!
> I have had a request to do a Miller's Elastic Stain. I have almost all of
> the stains and I do have all of the chemicals to do the stain but I have a
> question.
> One of the ingredients in the elastic stain is resorcin. I'm not a chemist
> so I have to ask this question, I have resorcinol in my chemical cabinet -
> is this the same thing as resorcin? I can find resorcinol in the Sigma
> cat. but not resorcin.
>
> Thanks!
> Andi Grantham
> .....................................................................
> : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
> : Sr. Research Specialist University of Arizona :
> : (office: AHSC 4212) P.O. Box 245044 :
> : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
> : (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
> :...................................................................:
> http://www.cba.arizona.edu/histology-lab.html
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 10
Date: Mon, 29 Jan 2007 14:19:22 -0700
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Subject: Re: [Histonet] Miller's Elastic Stain
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<4.3.2.7.2.20070129141822.00e1a5e0 <@t> algranth.inbox.email.arizona.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Thanks! Glad to hear that it is the same thing. Looks like I'm good to go.
Andi
At 01:33 PM 1/29/2007 -0700, Andrea Grantham wrote:
>Happy Monday!
>I have had a request to do a Miller's Elastic Stain. I have almost all of
>the stains and I do have all of the chemicals to do the stain but I have a
>question.
>One of the ingredients in the elastic stain is resorcin. I'm not a chemist
>so I have to ask this question, I have resorcinol in my chemical cabinet -
>is this the same thing as resorcin? I can find resorcinol in the Sigma
>cat. but not resorcin.
>
>Thanks!
>Andi Grantham
>.....................................................................
>: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
>: Sr. Research Specialist University of Arizona :
>: (office: AHSC 4212) P.O. Box 245044 :
>: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
>: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
>:...................................................................:
> http://www.cba.arizona.edu/histology-lab.html
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist University of Arizona :
: (office: AHSC 4212) P.O. Box 245044 :
: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
------------------------------
Message: 11
Date: Mon, 29 Jan 2007 15:20:56 -0600
From: Christina Thurby <christina.thurby <@t> bms.com>
Subject: [Histonet] Re: Anybody doing flow cytometry? What does Neat
mean?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <45BE6538.4090202 <@t> bms.com>
Content-Type: text/plain; charset="iso-8859-1"
Hello Histonet Folks!,
Is anyone out there performing flow cytometry that can tell me what a
suggested working dilution "Neat" means. I think it is the use of an
'undiluted' primary antibody but if someone familiar with this term can
please confirm that I'd be very appreciative.
Thanks,
Christina
------------------------------
Message: 12
Date: Mon, 29 Jan 2007 13:30:17 -0800 (PST)
From: Stasha McCollum <babistace52 <@t> yahoo.com>
Subject: [Histonet] ASCP Exam
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20070129213017.85649.qmail <@t> web35009.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Can anyone tell me if the practical portion of the certification exam has been remove? Ther were rumors floating about that change last year at this time. Also if anyone has taken it recently what is the percentage of IHC on the exam
Thanks
---------------------------------
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Message: 13
Date: Mon, 29 Jan 2007 13:44:41 -0800
From: GMartin <@t> marshallhospital.org
Subject: [Histonet] Processing debate
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <OFD4CDEC50.0465C64A-ON88257272.00764A56 <@t> com>
Content-Type: text/plain; charset=us-ascii
We presently process with a Tissue Tek Vip 2000. Our pathologist cut
the tissue in. the debate is this ... one side says that the cassettes
must be placed in a container of formalin to float at random before being
placed into the Vip 2000 basket.
The other side of the debate says that the tissue cassettes can be
placed in order directly into the basket that is submerged in formalin.
Can the group shine any light on this so far civil debate :)
Thanks
Gary Martin
El Dorado Pathology
California
------------------------------
Message: 14
Date: Mon, 29 Jan 2007 22:56:26 +0100
From: "nizar limaiem" <nizarlim <@t> gmail.com>
Subject: [Histonet] a question
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<92805bf10701291356j3e107923md8a75f1a0929f8a7 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Dear Doctor
I am student in biotechnology (Tunisia), I work on the developpement of the
Embryo
and Early Larva of a fish species dicentrarchus labrax L (it's egg is about
1mm in diameter).
I want to use methods of histology on fertilized eggs. Could some one offer
some suggestions on the experimental protocol
(fixation/processing/sectioning)
Thank you for your reply
limaiem nizar.
Student in marin biotechnology
University of biotechnology, Monastir, Tunisise
------------------------------
Message: 15
Date: Mon, 29 Jan 2007 17:08:03 -0500
From: "Kaye Ryan" <ryakay <@t> shands.ufl.edu>
Subject: Re: [Histonet] ASCP Exam
To: <histonet <@t> lists.utsouthwestern.edu>, <babistace52 <@t> yahoo.com>
Message-ID: <s5be2a09.047 <@t> GW-FS1.SHANDS.UFL.EDU>
Content-Type: text/plain; charset=US-ASCII
Hi,
Yes, the practical portion of the exam has been done away with. I had a
student that just took the exam. There are many more troubleshooting
questions now on all aspects of histology. My recommendations based on
what the student told me is that you need to really focus on
troubleshooting of special stains as well as fixation and cutting. As
far as I can tell, the study guide has not been updated and I don't know
if it will be.
Good luck,
Kaye Ryan
Kaye Ryan
Histology Manager/Educational Coordinator
Shands Rocky Point Laboratories
(352) 265-0111, 72093
>>> Stasha McCollum <babistace52 <@t> yahoo.com> 01/29/07 4:30 PM >>>
Can anyone tell me if the practical portion of the certification exam
has been remove? Ther were rumors floating about that change last year
at this time. Also if anyone has taken it recently what is the
percentage of IHC on the exam
Thanks
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Message: 16
Date: Mon, 29 Jan 2007 15:36:54 -0700
From: "Liz Chlipala" <liz <@t> premierlab.com>
Subject: [Histonet] human total IgG
To: "'Histonet'" <histonet <@t> pathology.swmed.edu>
Message-ID: <000001c743f5$f9f72280$0d00a8c0 <@t> domain.Premier>
Content-Type: text/plain; charset="us-ascii"
Hello everyone
Does anyone out there know of an antibody that will detect total IgG for
human tissue in paraffin sections. I have searched the web and have been
unable to come up with one that will detect total IgG so any help would be
appreciated.
Thanks in advance
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309
------------------------------
Message: 17
Date: Mon, 29 Jan 2007 17:29:30 -0500
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] Re: Anybody doing flow cytometry? What does
Neat mean?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4EBFF65383B74D49995298C4976D1D5E273C11 <@t> LSRIEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain; charset="iso-8859-1"
Yep, it means the same thing for antibodies that it means for scotch - undiluted - straight from the bottle.
> ----------
> From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Christina Thurby
> Sent: Monday, January 29, 2007 1:20 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Re: Anybody doing flow cytometry? What does Neat mean?
>
> <<File: ATT21946500.txt>>
>
> Hello Histonet Folks!,
> Is anyone out there performing flow cytometry that can tell me what a
> suggested working dilution "Neat" means. I think it is the use of an
> 'undiluted' primary antibody but if someone familiar with this term can
> please confirm that I'd be very appreciative.
> Thanks,
> Christina
>
>
>
------------------------------
Message: 18
Date: Mon, 29 Jan 2007 16:37:31 -0600
From: "Brown, Graham W" <graham.brown <@t> ttuhsc.edu>
Subject: [Histonet] MCP1 positive control
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <7F7DE44DEE269C4CB34BB8EADE0525EC61E472 <@t> BOWIE.ttuhsc.edu>
Content-Type: text/plain; charset="iso-8859-1"
Does anyone know of a positive control tissue for MCP1??
thanks
Graham Brown
Texas Tech University HSC
Garrison Institute on Aging
Dept. Neurology
Lubbock, TX
------------------------------
Message: 19
Date: Tue, 30 Jan 2007 09:19:15 +1000
From: Laurie Reilly <laurie.reilly <@t> jcu.edu.au>
Subject: Re: [Histonet] Mouse Testes Fixation and Processing
To: djemge <@t> aol.com, histonet <@t> lists.utsouthwestern.edu
Message-ID: <5.2.0.9.0.20070130090758.00bea1f0 <@t> mail.jcu.edu.au>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Donna and All,
Bouin's is the fixative of choice for testis, and my experience with
formalin fixation of testis tells me that your observations are correct.
A formalin-acetic acid-ethanol mix may be better. Maybe Davidson's fixative?
Another "trick of the trade" is to start the processing of testis (and
other very delicate tissues) in 30% Ethanol then 50%, 70%, 80%, 90%, 95%,
100%, 100% then 50:50 EtOH:Xylene, 2x Xylene 3x Paraffin.
For mouse tissues 15 minutes in each of these reagents would be enough.
Regards, Laurie.
At 11:18 AM 29/01/2007 -0500, djemge <@t> aol.com wrote:
>Hello All. I am having an ongoing problem with formalin fixed, paraffin
>embedded whole mouse testes (Adult mouse and Pups). I need to get this
>issue resolved and have consistent results for my investigators. They are
>using these testis for xgal, IF, IHC and In Situ. All my Bouin's fixed
>testes looks beautiful but the formalin fixed testes are horrible. Bouins
>is not suitable for much of what they are doing. I have tried several
>processing schedules and nothing seems to work. Sometimes the FFPE's seem
>very dry and shrunk, sometimes the cells look like they have big halos
>around them and you can't distinguish them enough for staging. This is
>frustrating! All other tissue seems to be fine except these and there is a
>big variability in how these testes turn out. They are NBF or cold 4%PFA
>PBS fixed for 24 hours. Typical processing schedule is either 30 minutes
>each station or 1 hour each station (formalin, 2x70%, 80%, 2x95%, 2x100%
>reagent alcohol, 2x Xylene, 3x Ameraffin Paraffin!
> (Cardinal cat# M7346-1A).
>
>Donna J. Emge, HT(ASCP)
>Northwestern University
>Lurie 7-220
>303 E. Superior Avenue
>Chicago, IL 60611
>312-503-2036
>d-emge <@t> northwestern.edu
>________________________________________________________________________
>Check out the new AOL. Most comprehensive set of free safety and security
>tools, free access to millions of high-quality videos from across the web,
>free AOL Mail and more.
>_______________________________________________
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>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Mr.Laurie Reilly
School of Veterinary and Biomedical Sciences
James Cook University
Townsville. 4811
Australia.
Phone 07 4781 4468
Fax 07 4779 1526
------------------------------
Message: 20
Date: Mon, 29 Jan 2007 15:34:38 -0800 (PST)
From: Robert Chiovetti <rchiovetti <@t> yahoo.com>
Subject: Re: [Histonet] Immunogold question
To: Danielle Crippen <dcrippen <@t> buckinstitute.org>,
histonet <@t> lists.utsouthwestern.edu
Message-ID: <825913.37628.qm <@t> web58903.mail.re1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Hi Danielle,
Just a guess here, but the higher pH may help reduce nonspecific background staining.
I worked in an EM lab that occasionally played with pH and salt concentrations for immunogold labeling when it was necessary. I know for sure that high salt washes after the primary and secondary Ab's, followed by a return to normal TBS, does a super job of getting rid of background. I'm not so certain about the pH. I only used it a couple of times and I really couldn't tell much difference...
Bob
Robert (Bob) Chiovetti, Ph.D.
Southwest Precision Instruments
Arizona's Microscopy Resource
132 North Elster Drive
Tucson, AZ 85710-3212
Tel./Fax 520-546-4986
Member, Arizona Small Business Association
(www.asba.com)
----- Original Message ----
From: Danielle Crippen <dcrippen <@t> buckinstitute.org>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Monday, January 29, 2007 10:27:46 AM
Subject: [Histonet] Immunogold question
Dear EM experts,
I've been using the following protocol for immunogold labeling for the past couple years with good success. This morning one of our users has inquired as to why the secondary is incubated at a higher pH than the primary. I do not know the answer...do any of you???
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Message: 21
Date: Mon, 29 Jan 2007 16:06:53 -0800
From: GMartin <@t> marshallhospital.org
Subject: [Histonet] Follow up
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <OF7FF673CC.A329343D-ON88257272.00818324 <@t> com>
Content-Type: text/plain; charset=us-ascii
I'm new to the list and in an effort to keep the list informed I want to
thank you for the response to my two questions.
1) concerning Xylene substitutes ... I received a good suggestion on a
product FORMULA 83. And I will as suggested ... report back on my success
or failures of my transition from Xylene to a substitute.
2) Processing cassettes ... the debate was weather to put them directly
into the processing basket or let them float randomly in formalin then put
them in the basket. The standard seems to be to load them directly from
the grossing table into the processing basket.
Thank you all very much
Gary Martin
El Dorado Pathology
------------------------------
Message: 22
Date: Mon, 29 Jan 2007 16:31:31 -0800
From: "Tarango, Mark" <mtarango <@t> nvcancer.org>
Subject: RE: [Histonet] Re: Anybody doing flow cytometry? What does
Neat mean?
To: "Christina Thurby" <christina.thurby <@t> bms.com>,
histonet <@t> lists.utsouthwestern.edu
Message-ID:
<5AEC610C1CE02945BD63A395BA763EDE1F9951 <@t> NVCIEXCH02.NVCI.org>
Content-Type: text/plain; charset=us-ascii
You're right. It means using it straight.
Mark Adam Tarango HT(ASCP)
Histology/Immunohistochemistry Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV 89135
mtarango <@t> nvcancer.org
Direct Line (702) 822-5112
Fax (702) 939-7663
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Christina Thurby
Sent: Monday, January 29, 2007 1:21 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Anybody doing flow cytometry? What does Neat
mean?
Hello Histonet Folks!,
Is anyone out there performing flow cytometry that can tell me what a
suggested working dilution "Neat" means. I think it is the use of an
'undiluted' primary antibody but if someone familiar with this term can
please confirm that I'd be very appreciative.
Thanks,
Christina
"EMF <nvcancer.org>" made the following annotations.
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Message: 23
Date: Mon, 29 Jan 2007 18:45:11 -0800 (PST)
From: MICHELLE SEAGLE <mrsseagle <@t> yahoo.com>
Subject: [Histonet] ASCP EXAM
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <831462.68750.qm <@t> web51812.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
I just took HT exam this past december and I dont remember having any IHC related questions on the exam. You just need to know the freida carson book backwards and forwards because they pick the starngest things out of that book that you would not know unless you have just practically memorized the book!! Don't wast your time or money on the ASCP practical exam questions, what a waste. As far as the practical part I am not sure but I believe It has been Removed from the requirements since everthing is so automated now.
Good Luck on your Exam
Michelle Seagle HT (ASCP)
Rutherford Hospital
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Message: 24
Date: Tue, 30 Jan 2007 06:53:19 +0100
From: "Jakub Otahal" <jotahal <@t> epilepsy.biomed.cas.cz>
Subject: Re: [Histonet] Hypoxia
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20070130055319.9dbd9a05 <@t> epilepsy.biomed.cas.cz>
Content-Type: text/plain; charset="us-ascii"
Dear Gamal,
have a look at chemicon site (www.chemicon.com) and search for hypoxyprobe. You will have staining kit as well good productsheet describing principles of this method and staining procedure. there is good reference list also. It is good to start with......
Jakub
*****************************************
Jakub Otahal MD,PhD
Department of Developmental Epileptology
Institute of Physiology
Czech Academy of Sciences
Videnska 1083
14220 Prague 4
jotahal <@t> epilepsy.biomed.cas.cz
http://epilepsy.biomed.cas.cz
tel: +420 241062495
*****************************************
_____
From: Gamal Akabani [mailto:gamal.akabani <@t> gmail.com]
To: Histonet ((E-mail)) [mailto:histonet <@t> lists.utsouthwestern.edu]
Sent: Mon, 29 Jan 2007 21:40:39 +0100
Subject: [Histonet] Hypoxia
Dear friends,
I am want to process some histological samples from glioma patients
in order to observe the level or grade of hypoxia on them, if any.
I am not a guru. Any help is appreciated on the methods or a
specific antibody used to visualize it. Thanks in advanced.
Gamal
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------------------------------
Message: 25
Date: Tue, 30 Jan 2007 08:37:27 -0500
From: "Houston, Ronald" <HoustonR <@t> chi.osu.edu>
Subject: [Histonet] RUO's and diagnostic IHC
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<979FF5962E234F45B06CF0DB7C1AABB20D77CEFF <@t> chi2k3ms01.columbuschildrens.net>
Content-Type: text/plain; charset="us-ascii"
Here is a reply I received yesterday from Dr Steve Gutman, Director, In
Vitro Diagnostics, FDA, regarding the current status of using RUOs in
the clinical lab. He did add the disclaimer that these were his views
and did not represent official FDA policy; and you thought it was
confusing before!!!!!
Don't shoot the messenger.........
"-----Original Message-----
From: Gutman, Steve [mailto:steve.gutman <@t> fda.hhs.gov]
Sent: Monday, January 29, 2007 2:52 PM
To: Houston, Ronald
Subject: RE: RUO's and ASR reagents in diagnostics - FDA disclaimer
I have had informal discussions with CAP but FDA has not taken a formal
position on use of RUOs to support home brews. In general, we think
this is a dreadful choice and clearly a tactic of last resort. There
are no regulatory requirements for RUO devices. So the laboratory takes
full responsibility for ensuring quality of production of the RUO
material being made over time. This seems to me to put laboratories at
considerable liability risk since the general controls applied to ASRs
don't work here.
As we try to review our regulatory policy in this area, it is certainly
one we should and will try to clarify.
Thanks.
Steve
________________________________
From: Houston, Ronald [mailto:HoustonR <@t> chi.osu.edu]
Sent: Monday, January 29, 2007 10:23 AM
To: Gutman, Steve
Subject: RUO's and ASR reagents in diagnostics - FDA disclaimer
Importance: High
Dr Gutman,
There is tremendous confusion in laboratory circles regarding the FDA
disclaimer for ASR antibodies and whether or not this same disclaimer
can be used for RUO antibodies, particularly in light of CAP's change in
interpretation of the guidelines for IHC testing:
"Antibodies, nucleic acid sequences, etc., labeled "Research Use Only"
(RUO) purchased from commercial sources may be used in home brew test
only if the laboratory has made a reasonable effort to search for IVD or
ASR class reagents. The results of that failed search should be
documented by the laboratory director.
The laboratory must establish or verify the performance characteristics
of tests using Class I ASR's and RUO's in accordance with the Method
Performance Specifications section of the Laboratory General checklist."
Exactly what is the current stance of the FDA, and have any discussions
taken place between the FDA and CAP regarding their interpretation? Can
results of RUO testing be dictated into a diagnostic report and can the
test be billed?
Obviously, antibody manufacturers mandate that their RUO antibodies
cannot be used for diagnostic purposes, but they are as confused as the
laboratorians.
Thank you for taking the time to clarify this very confusing situation.
Sincerely
Ronnie Houston, MS, HT(ASCP)QIHC
Anatomic Pathology Manager
Columbus Children's Hospital
700 Children's Drive
Columbus, OH 43205
(614) 722 5465
houstonr <@t> chi.osu.edu <mailto:houstonr <@t> chi.osu.edu> "
Ronnie Houston, MS, HT(ASCP)QIHC
Anatomic Pathology Manager
Columbus Children's Hospital
700 Children's Drive
Columbus, OH 43205
(614) 722 5465
houstonr <@t> chi.osu.edu <mailto:houstonr <@t> chi.osu.edu>
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Message: 26
Date: Tue, 30 Jan 2007 07:02:53 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Processing debate
To: GMartin <@t> marshallhospital.org, histonet <@t> lists.utsouthwestern.edu
Message-ID: <189401.17374.qm <@t> web61213.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Fear the word "random", in histology is almost synonym with "caos".There is absolutely no logic or benefit from placing the cassettes at random.
If you use a cassetting log the sequencial and orderly manner of placing the cassettes following the log order is the way of doing things.
You can always know where your cassettes are and at the embedding step you can follow the same sequency and easily determine if all the cassettes were processed and embeded.
It is absolutely illogical to place anything at random (unless you are selecting something for an experimental design that requires that type of selection!).
The only "logical" explanation to your initial question is that it stems from the pathologist's lazyness that prefers to throw the cassettes in the container and letting the histotech to try to organize what should not have been disorganized in the first place.
René J.
GMartin <@t> marshallhospital.org wrote:
We presently process with a Tissue Tek Vip 2000. Our pathologist cut
the tissue in. the debate is this ... one side says that the cassettes
must be placed in a container of formalin to float at random before being
placed into the Vip 2000 basket.
The other side of the debate says that the tissue cassettes can be
placed in order directly into the basket that is submerged in formalin.
Can the group shine any light on this so far civil debate :)
Thanks
Gary Martin
El Dorado Pathology
California
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