[Histonet] Re: a basic question about immunohistochemistry (IHC ON
DEAD RAT)
Jamie E Erickson
jamie.erickson <@t> abbott.com
Mon Nov 30 07:55:03 CST 2009
Dear Salim,
I have done lots of IHC on mouse and rat tissues ie brain and trying to
make correlations on viable tissue is hard enough. If you do preform the
IHC it may confound your data one way or the other and leave you with more
questions then answers.
My recommendation would be to leave this animal out of the analysis.
Autolysis can be difficult to read through with IHC in the brain.
Ultimately it is up to you but if this animal shows more or less staining
and skews your average in that group you will be left with the nagging
question " was it the autolysis that made the difference"
My 2 cents...good luck..
Jamie E Erickson
Scientist II, M.S. HTL (ASCP)
Discovery Safety, Metabolism & Pharmacokinetics
Abbott Laboratories
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Histonet Digest, Vol 72, Issue 33
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Today's Topics:
1. RE: a basic question about immunohistochemistry (Thomas Jasper)
2. AW: [Histonet] Biopsy fixation (Nathan Jentsch)
3. RE: SPAM-LOW: RE: [Histonet] a basic question about
immunohistochemistry (Patsy Ruegg)
4. AW: [Histonet] Biopsy fixation (Gudrun Lang)
5. Cassette labelling (Anne van Binsbergen)
6. Re: a basic question about immunohistochemistry (Lesley Weston)
----------------------------------------------------------------------
Message: 1
Date: Sat, 28 Nov 2009 10:17:11 -0800
From: "Thomas Jasper" <tjasper <@t> copc.net>
Subject: RE: [Histonet] a basic question about immunohistochemistry
To: "Salim Yalcin Inan" <syinan <@t> ucalgary.ca>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<90354A475B420441B2A0396E5008D4965E30A8 <@t> copc-sbs.COPC.local>
Content-Type: text/plain; charset="us-ascii"
Dear Salim,
As you have been informed, doing immunohistochemistry is possible on
this tissue. After all it's possible to do IHC on any tissue whether
the conditions you want to test under are ideal or not.
Being chastised on this list and calling your work "bad science" is
totally out of line and certainly does not help you out. I think some
people would do well to reserve judgment, particularly when there's no
way they can fully understand what's going on with your project. Having
worked in research myself, I completely understand that animals will
die, at the most inconvenient times, during a study.
First of all you should incorporate the data about the animal dying into
your study notes. Secondly, there's no harm in running the IHC on this
animal's tissue. You can use the results comparatively with results
from some perfused tissue later on.
I don't know Salim, some people might call it damage control, or making
the best of a less than ideal situation. Again, I don't know exactly
what you're working on but it seems there's information worth gathering
despite the circumstances. I also understand that it's probably next to
impossible to carry out experiments and research alone. Having reliable
staff assist you is not unusual either.
Good luck to you,
Tom Jasper
Thomas Jasper HT (ASCP) BAS
Histology Supervisor
Central Oregon Regional Pathology Services
Bend, Oregon 97701
541/693-2677
tjasper <@t> copc.net
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Salim
Yalcin Inan
Sent: Friday, November 27, 2009 2:44 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] a basic question about immunohistochemistry
Dear All,
Because I am new in immunohistochemistry, I have a basic question about
it.
What if your rat dies in the evening or in the weekend, which you are
doing a chronic experiment and need to collect brain tissue for
immunohistochemistry? And let's say, the staff did not noticed it to
inform you on time. Several hours passed since your rat died. There is
no way to do perfusion. Is it still possible to do immunohistochemistry?
Thank you very much in advance.
Best regards,
Salim Yalcin Inan, Ph.D.
(post-doctoral fellow)
Department of Clinical Neurosciences
University of Calgary
syinan <@t> ucalgary.ca
_______________________________________________
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------------------------------
Message: 2
Date: Sat, 28 Nov 2009 17:35:31 -0500
From: Nathan Jentsch <njblademaster <@t> gmail.com>
Subject: AW: [Histonet] Biopsy fixation
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<5a7745af0911281435j339ff9d3ja8d7eff31e93167b <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Gudrun
I work in a dermatopathology lab, and we don't see any issues leaving our
specimens in 10% NBF for days at a time. We don't work on the weekends,
so
any specimens that are grossed remain on our processors in NBF from Friday
at 8:00 PM until about 5:30 PM on Sunday. I also agree with Kris that 4
hours of fixation is not long enough to ensure proper fixation unless they
are shave biopsies.
Nate
------------------------------
Message: 3
Date: Sat, 28 Nov 2009 15:55:24 -0700
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: SPAM-LOW: RE: [Histonet] a basic question about
immunohistochemistry
To: "'Thomas Jasper'" <tjasper <@t> copc.net>, "'Salim Yalcin
Inan'"
<syinan <@t> ucalgary.ca>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <A13FCBAEA5394327948047717FE837B5 <@t> prueggihctechlt>
Content-Type: text/plain; charset="us-ascii"
Of course you can always do IHC on your rat that died, you just have to
note
that that rat died and was not profuse fixed so that the review of the
results will take that in consideration, hopefully you have another rat
that
besides dyeing and not getting profuse fixed had all other conditions the
mostly the same so you could use the properly treated rat as a standard to
compare the dead rat too.
Good luck and do keep asking for help at this forum, most of us will offer
you our experience without judging your science.
Regards,
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net
www.ihcrg.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Thomas
Jasper
Sent: Saturday, November 28, 2009 11:17 AM
To: Salim Yalcin Inan
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: RE: [Histonet] a basic question about
immunohistochemistry
Dear Salim,
As you have been informed, doing immunohistochemistry is possible on
this tissue. After all it's possible to do IHC on any tissue whether
the conditions you want to test under are ideal or not.
Being chastised on this list and calling your work "bad science" is
totally out of line and certainly does not help you out. I think some
people would do well to reserve judgment, particularly when there's no
way they can fully understand what's going on with your project. Having
worked in research myself, I completely understand that animals will
die, at the most inconvenient times, during a study.
First of all you should incorporate the data about the animal dying into
your study notes. Secondly, there's no harm in running the IHC on this
animal's tissue. You can use the results comparatively with results
from some perfused tissue later on.
I don't know Salim, some people might call it damage control, or making
the best of a less than ideal situation. Again, I don't know exactly
what you're working on but it seems there's information worth gathering
despite the circumstances. I also understand that it's probably next to
impossible to carry out experiments and research alone. Having reliable
staff assist you is not unusual either.
Good luck to you,
Tom Jasper
Thomas Jasper HT (ASCP) BAS
Histology Supervisor
Central Oregon Regional Pathology Services
Bend, Oregon 97701
541/693-2677
tjasper <@t> copc.net
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Salim
Yalcin Inan
Sent: Friday, November 27, 2009 2:44 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] a basic question about immunohistochemistry
Dear All,
Because I am new in immunohistochemistry, I have a basic question about
it.
What if your rat dies in the evening or in the weekend, which you are
doing a chronic experiment and need to collect brain tissue for
immunohistochemistry? And let's say, the staff did not noticed it to
inform you on time. Several hours passed since your rat died. There is
no way to do perfusion. Is it still possible to do immunohistochemistry?
Thank you very much in advance.
Best regards,
Salim Yalcin Inan, Ph.D.
(post-doctoral fellow)
Department of Clinical Neurosciences
University of Calgary
syinan <@t> ucalgary.ca
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 4
Date: Sun, 29 Nov 2009 11:18:52 +0100
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] Biopsy fixation
To: "'Nathan Jentsch'" <njblademaster <@t> gmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <355415E1E34E48EE854F2D0A668F81E4 <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
Nate,
I think you have confused the persons. I'm the one, who recommended longer
fixation and Kris had made the original question.
Regards
Gudrun
-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Nathan
Jentsch
Gesendet: Samstag, 28. November 2009 23:36
An: histonet <@t> lists.utsouthwestern.edu
Betreff: AW: [Histonet] Biopsy fixation
Gudrun
I work in a dermatopathology lab, and we don't see any issues leaving our
specimens in 10% NBF for days at a time. We don't work on the weekends,
so
any specimens that are grossed remain on our processors in NBF from Friday
at 8:00 PM until about 5:30 PM on Sunday. I also agree with Kris that 4
hours of fixation is not long enough to ensure proper fixation unless they
are shave biopsies.
Nate
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Sun, 29 Nov 2009 16:24:20 +0400
From: Anne van Binsbergen <annigyg <@t> gmail.com>
Subject: [Histonet] Cassette labelling
To: "histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>,
histonet-request <@t> lists.utsouthwestern.edu
Message-ID:
<f8332fbe0911290424w2c0f8a9dx1b0214168cc6a5d3 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi Histonetters
Who out there has a Sakura iDent?
I have just taken delivery of a brand new machine and I am having serious
issues with the quality of the print on the cassette face - the ink rubs
off
with xylene!!!
Yes, I leave the print to dry before I handle the cassettes
No, I dont use Tissue-Tek xylene substitute - my xylene is made by Fisher
I would love to know if anyone else has had similar issues
Yes, Rene, this is the diehard number 1 Sakura fan having issues with a
Sakura product!!!
hopefully yours (pun intended)
AbuDhabiAnnie
--
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
------------------------------
Message: 6
Date: Sun, 29 Nov 2009 06:22:57 -0800
From: Lesley Weston <leswes <@t> shaw.ca>
Subject: Re: [Histonet] a basic question about immunohistochemistry
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <BF6C7B78-0305-48E3-A6E2-584BE0E2C6EF <@t> shaw.ca>
Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed
Just about all biology research institutions have an animal unit with
qualified technicians to look after the animals, rather than someone
qualified in other areas; however the technicians are subject to
human failings such as going home at night. I've never heard of a one-
person research team, and I don't think it would be all that effective.
The tissue from the animal that died at the wrong time is
not
comparable to perfused tissue, but as Tom says, it might still be
worth processing. If nothing else, it will show how much difference
perfusion makes.
Lesley Weston.
On 28-Nov-09, at 10:17 AM, Thomas Jasper wrote:
> Dear Salim,
>
> As you have been informed, doing immunohistochemistry is possible on
> this tissue. After all it's possible to do IHC on any tissue whether
> the conditions you want to test under are ideal or not.
>
> Being chastised on this list and calling your work "bad science" is
> totally out of line and certainly does not help you out. I think some
> people would do well to reserve judgment, particularly when there's no
> way they can fully understand what's going on with your project.
> Having
> worked in research myself, I completely understand that animals will
> die, at the most inconvenient times, during a study.
>
> First of all you should incorporate the data about the animal dying
> into
> your study notes. Secondly, there's no harm in running the IHC on
> this
> animal's tissue. You can use the results comparatively with results
> from some perfused tissue later on.
>
> I don't know Salim, some people might call it damage control, or
> making
> the best of a less than ideal situation. Again, I don't know exactly
> what you're working on but it seems there's information worth
> gathering
> despite the circumstances. I also understand that it's probably
> next to
> impossible to carry out experiments and research alone. Having
> reliable
> staff assist you is not unusual either.
>
> Good luck to you,
>
> Tom Jasper
>
>
> Thomas Jasper HT (ASCP) BAS
> Histology Supervisor
> Central Oregon Regional Pathology Services
> Bend, Oregon 97701
> 541/693-2677
> tjasper <@t> copc.net
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Salim
> Yalcin Inan
> Sent: Friday, November 27, 2009 2:44 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] a basic question about immunohistochemistry
>
> Dear All,
>
> Because I am new in immunohistochemistry, I have a basic question
> about
> it.
> What if your rat dies in the evening or in the weekend, which you are
> doing a chronic experiment and need to collect brain tissue for
> immunohistochemistry? And let's say, the staff did not noticed it to
> inform you on time. Several hours passed since your rat died. There is
> no way to do perfusion. Is it still possible to do
> immunohistochemistry?
> Thank you very much in advance.
>
> Best regards,
>
> Salim Yalcin Inan, Ph.D.
> (post-doctoral fellow)
> Department of Clinical Neurosciences
> University of Calgary
> syinan <@t> ucalgary.ca
>
>
------------------------------
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