[Histonet] RE: Polymer based IHC reagent for mouse antibodies on
rat tissue
Wynn, Carmen
Carmen.Wynn <@t> us.astellas.com
Wed Oct 17 09:51:01 CDT 2007
Hello Histonetters,
I have had great success with the Biocare HRP polymer product line. I
have used the various kits: Mouse Primary antibody on Rat tissue, Rat
Primary Antibody on Mouse tissue, Rabbit Primary on Rodent tissue (for
both mouse and rat), and Goat Primary antibody on Human, Mouse, and Rat
tissue. All work great with no background and the protocols are very
quick. Just don't forget to rinse in water prior to the substrate
chromagen application...VERY IMPORTANT!! I use Romulin AEC at double
strength for chromagen. They also offer all these reagents in the AP
form as well. Good Luck!
Carmen Wynn, M.S., Senior Scientist
Astellas Research Institute of America, LLC. (ARIA)
Illinois Science and Technology Park
8045 Lamon Ave
Skokie, IL 60077
Direct: 847-933-7419
Main: 847-933-7400
Fax: 847-933-7401
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Sent: Saturday, October 13, 2007 12:07 PM
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Subject: Histonet Digest, Vol 47, Issue 15
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Today's Topics:
1. Re: Teratogens (Rene J Buesa)
2. Re: Defrosting Cryostat (Rene J Buesa)
3. proventricular dilatation images? (I-sanna Gibbons)
4. Re: proventricular dilatation images? (Cheryl Cross)
5. Re: proventricular dilatation images? (I-sanna Gibbons)
6. Keeping frozen mouse aorta on slides (Randolph-Habecker, Julie)
7. polymer based IHC reagent for mouse antibodies on rat tissue
(Liz Chlipala)
8. Re: Keeping frozen mouse aorta on slides (Rene J Buesa)
9. Keeping frozen mouse aorta on slides (Paula Pierce)
10. polymer based IHC reagent for mouse antibodies on rat tissue
(Paula Pierce)
----------------------------------------------------------------------
Message: 1
Date: Fri, 12 Oct 2007 10:23:30 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Teratogens
To: Diana McCaig <dmccaig <@t> ckha.on.ca>,
histonet <@t> lists.utsouthwestern.edu
Message-ID: <823783.2629.qm <@t> web61214.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Diana:
1- teratogen, mutagen or any such an indication in the MSDS should be
considered as warnings for the label
2- at any concentration
3- all pregnant employees should (and deserve) being assigned low risk
activities, even when 31% of USA histology labs do not practice this
prohibition
4- you could Google "Teratogen and Mutagen"
Ren J.
Diana McCaig <dmccaig <@t> ckha.on.ca> wrote:
Hi
I was wondering if anyone specifically labels these chemicals in your
lab to alert any techs of child producing age.
Also, how do you establish this criteria, some could be in the
concentrated form or powder state, but do they still qualify in a dilute
state?
Is this determined strictly by the MSDS and based on the word teratogen
or do you also consider terminology as mutagen, reproductive effector as
well?
If protective equipment and proper ventilation is available, are
pregnant staff exempt for doing any activity or stain that uses these
solutions for the full term of their pregnancy and "I want to become
pregnant" stages?
And finally, can another provide a list of the chemicals they class as
teratogens.
thanks
Diana McCaig
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Message: 2
Date: Fri, 12 Oct 2007 10:24:59 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Defrosting Cryostat
To: "Piche-Grocki, Jessica" <jpiche-grocki <@t> wtbyhosp.org>,
histonet <@t> lists.utsouthwestern.edu
Message-ID: <709197.83606.qm <@t> web61222.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Cryostats should be defrosted daily, near midnight.
Ren J.
"Piche-Grocki, Jessica" <jpiche-grocki <@t> wtbyhosp.org> wrote:
Hi,
How often should we defrost our Cryostat? I have set it up for once a
week. Should it be more often?
Thanks,
Jessica Piche-Grocki, HT(ASCP)
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Message: 3
Date: Fri, 12 Oct 2007 11:04:36 -0700 (PDT)
From: I-sanna Gibbons <trinimaican2501 <@t> yahoo.com>
Subject: [Histonet] proventricular dilatation images?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <361313.20681.qm <@t> web50310.mail.re2.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Hi all,
Does anyone have histological images of proventricular dilatation and
also images of the normal for comparison?
Thanks
I-sanna
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Message: 4
Date: Fri, 12 Oct 2007 14:27:07 -0400
From: Cheryl Cross <ccross6032 <@t> aol.com>
Subject: Re: [Histonet] proventricular dilatation images?
To: I-sanna Gibbons <trinimaican2501 <@t> yahoo.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <05462208-2622-43F5-95C0-52961EF6A3A0 <@t> aol.com>
Content-Type: text/plain; charset=US-ASCII; delsp=yes;
format=flowed
Hi I-sanna -
I don't have any images readily available - there is a nice little
review with some histopathology here, however:
http://www.vet.uga.edu/vpp/ivcvm/1998/gregory/index.php
What I can tell you is that in many cases can be a very very subtle
disease - often there are just a few lymphocytes around the nerve
ganglia - very very subtle. if you look at just a normal
proventriculus histology book - an animal with really remarkable with
have a thinned, stretched out proventriculus, but the histology can
look very similar. It's easier to diagnose grossly (in my experience
anyway), because the amount of inflammation histopathologically
doesn't always match up with the degree of dilatation (ie animals
with a marked dilation can have minimal inflammation).
make sense?
CC
Cheryl Cross, DVM, Dipl. ACVP
Researcher
University Corporation for Atmospheric Research
College of Veterinary Medicine
University of Tennessee Department of Pathology
2407 River Drive, Room A201
Knoxville, TN 37996-4542
(423) 967-2724
fax: 865-974-5616
ccross <@t> ucar.edu
On Oct 12, 2007, at 2:04 PM, I-sanna Gibbons wrote:
> Hi all,
>
> Does anyone have histological images of proventricular dilatation
> and also images of the normal for comparison?
>
> Thanks
> I-sanna
>
>
>
> ______________________________________________________________________
> ______________
> Moody friends. Drama queens. Your life? Nope! - their life, your
> story. Play Sims Stories at Yahoo! Games.
> http://sims.yahoo.com/
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Fri, 12 Oct 2007 12:21:39 -0700 (PDT)
From: I-sanna Gibbons <trinimaican2501 <@t> yahoo.com>
Subject: Re: [Histonet] proventricular dilatation images?
To: Cheryl Cross <ccross6032 <@t> aol.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <24335.52735.qm <@t> web50301.mail.re2.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Cheryl
Yes it does! Thanks. Will check the website
I-sanna
----- Original Message ----
From: Cheryl Cross <ccross6032 <@t> aol.com>
To: I-sanna Gibbons <trinimaican2501 <@t> yahoo.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Sent: Friday, October 12, 2007 3:27:07 PM
Subject: Re: [Histonet] proventricular dilatation images?
Hi I-sanna -
I don't have any images readily available - there is a nice little
review with some histopathology here, however:
http://www.vet.uga.edu/vpp/ivcvm/1998/gregory/index.php
What I can tell you is that in many cases can be a very very subtle
disease - often there are just a few lymphocytes around the nerve
ganglia - very very subtle. if you look at just a normal proventriculus
histology book - an animal with really remarkable with have a thinned,
stretched out proventriculus, but the histology can look very similar.
It's easier to diagnose grossly (in my experience anyway), because the
amount of inflammation histopathologically doesn't always match up with
the degree of dilatation (ie animals with a marked dilation can have
minimal inflammation).
make sense?
CC
Cheryl Cross, DVM, Dipl. ACVP
Researcher
University Corporation for Atmospheric Research
College of Veterinary Medicine
University of Tennessee Department of Pathology
2407 River Drive, Room A201
Knoxville, TN 37996-4542
(423) 967-2724
fax: 865-974-5616
ccross <@t> ucar.edu
On Oct 12, 2007, at 2:04 PM, I-sanna Gibbons wrote:
Hi all,
Does anyone have histological images of proventricular dilatation and
also images of the normal for comparison?
Thanks
I-sanna
________________________________________________________________________
____________
Moody friends. Drama queens. Your life? Nope! - their life, your story.
Play Sims Stories at Yahoo! Games.
http://sims.yahoo.com/
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Message: 6
Date: Fri, 12 Oct 2007 15:18:40 -0700
From: "Randolph-Habecker, Julie" <jhabecke <@t> fhcrc.org>
Subject: [Histonet] Keeping frozen mouse aorta on slides
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <040346FA7309BD439C327F97D4C4D69B13F127 <@t> ISIS.fhcrc.org>
Content-Type: text/plain; charset="us-ascii"
Folks,
I need your suggestions! I am working with an investigator who wants my
lab to stain unfixed, OCT embedded mouse aortas. Unfortunately, they
have already cut all of the tissue so we do not have a choice of slides.
They were cut at 6 microns on Star frost slides and stored at -80C for a
few months. When we received them, we took a few slides out, let them
air dry at room temperature for 24 hours, put them in a slides rack,
soaked them in room temperature PBS for 5 minutes and fixed them with
10% NBF for 10 minutes. These steps are done without agitation. This
procedure is compatible with our staining protocol and normally works
fine when we use Super frost or Gold Plus slides from Erie. However, in
this case, the tissue is floating off the slides in both the PBS and the
NBF stage. I have also tried to do these steps with the slides flat and
get the same results.
Any suggestion on ways to "glue" already cut frozen sections to a
charged slide in ways that are compatible with x-gal and IHC staining?
Thanks in advance for your help!!!!
Julie
Julie Randolph-Habecker, Ph.D.
Staff Scientist - Director
Experimental Histopathology Shared Resource
Fred Hutchinson Cancer Research Center
1100 Fairview Ave, N. M5-A803
Seattle WA 98109-1024
206-667-6119
jhabecke <@t> fhcrc.org
------------------------------
Message: 7
Date: Fri, 12 Oct 2007 16:28:14 -0600
From: "Liz Chlipala" <liz <@t> premierlab.com>
Subject: [Histonet] polymer based IHC reagent for mouse antibodies on
rat tissue
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<EE33BE5C905A3046A7FF8F58A64C8E4B0507E8 <@t> server.PremierLab.local>
Content-Type: text/plain; charset="us-ascii"
Hello all
Is anyone out there know of a polymer based reagent that will work for
mouse antibodies on rat tissues?
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 <http://www.premierlab.com/>
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309
------------------------------
Message: 8
Date: Sat, 13 Oct 2007 06:46:17 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Keeping frozen mouse aorta on slides
To: "Randolph-Habecker, Julie" <jhabecke <@t> fhcrc.org>,
histonet <@t> lists.utsouthwestern.edu
Message-ID: <260570.30359.qm <@t> web61212.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Julie:
The collagen nature of aortas make them more difficult to adhere.
I think that you will need a "mechanical barrier" to restrain the
sections. Try doing the whole procedure with the slides flat and the
sections covered by a very fine plastic mesh that will hold the sections
down by its weight. The mesh will allow all the reagents to act.
The only problem will be when lifting the mesh that the sections may
stick to it.
The procedure could also be done through a piece of filter paper, but
it is more likely to peel the sections than with the mesh.
Just a thought, you may try 1 or 2 sections to see what happens.
Ren J.
"Randolph-Habecker, Julie" <jhabecke <@t> fhcrc.org> wrote:
Folks,
I need your suggestions! I am working with an investigator who wants my
lab to stain unfixed, OCT embedded mouse aortas. Unfortunately, they
have already cut all of the tissue so we do not have a choice of slides.
They were cut at 6 microns on Star frost slides and stored at -80C for a
few months. When we received them, we took a few slides out, let them
air dry at room temperature for 24 hours, put them in a slides rack,
soaked them in room temperature PBS for 5 minutes and fixed them with
10% NBF for 10 minutes. These steps are done without agitation. This
procedure is compatible with our staining protocol and normally works
fine when we use Super frost or Gold Plus slides from Erie. However, in
this case, the tissue is floating off the slides in both the PBS and the
NBF stage. I have also tried to do these steps with the slides flat and
get the same results.
Any suggestion on ways to "glue" already cut frozen sections to a
charged slide in ways that are compatible with x-gal and IHC staining?
Thanks in advance for your help!!!!
Julie
Julie Randolph-Habecker, Ph.D.
Staff Scientist - Director
Experimental Histopathology Shared Resource
Fred Hutchinson Cancer Research Center
1100 Fairview Ave, N. M5-A803
Seattle WA 98109-1024
206-667-6119
jhabecke <@t> fhcrc.org
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Message: 9
Date: Sat, 13 Oct 2007 07:14:36 -0700 (PDT)
From: Paula Pierce <contact <@t> excaliburpathology.com>
Subject: [Histonet] Keeping frozen mouse aorta on slides
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <220369.5150.qm <@t> web50102.mail.re2.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Try going right to formalin after air drying, instead of PBS.
Paula Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
630 N. Broadway
Moore, OK 73160
405-570-6679
405-759-3953
contact <@t> excaliburpathology.com
------------------------------
Message: 10
Date: Sat, 13 Oct 2007 07:16:16 -0700 (PDT)
From: Paula Pierce <contact <@t> excaliburpathology.com>
Subject: [Histonet] polymer based IHC reagent for mouse antibodies on
rat tissue
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <122947.8101.qm <@t> web50106.mail.re2.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Mouse ImmPress from Vector Labs.
Paula Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
630 N. Broadway
Moore, OK 73160
405-570-6679
405-759-3953
contact <@t> excaliburpathology.com
------------------------------
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