[Histonet] Softening animal tissue
James Watson
jwatson <@t> gnf.org
Mon Feb 12 10:35:47 CST 2007
Diane,
For years I have been using 5% glycerin in my absolute alcohol on the
tissue processor for animal tissue here is a the program for mouse
tissue.
Mouse Tissue Processing Schedule
Station Reagent Temperature Vacuum Time Drain
Time Agitate
1 70% ETOH Ambient OFF
0:30 30 Off
2 80% ETOH Ambient Vacuum 0:30
30 Stirred
3 95% ETOH Ambient Vacuum 0:30
30 Stirred
4 95% ETOH Ambient Vacuum 0:45
45 T&S
5 5% Glycerin in100% ETOH Ambient Vacuum 0:30 30
Stirred
6 5% Glycerin in 100% ETOH Ambient Vacuum 0:30 30
Stirred
7 5% Glycerin in 100% ETOH Ambient V&P 0:45
100 T&S
8 Xylene Ambient Vacuum 0:30 45
Stirred
9 Xylene Ambient Vacuum 0:30 45
Stirred
10 Xylene Ambient V&P 0:45
120 T&S
11 Paraffin 60 V&P
0:30 120 Stirred
12 Paraffin 60 V&P
0:30 120 Stirred
13 Paraffin 60 V&P
0:45 120 Stirred
14 Paraffin 60 V&P
0:45 120 Stirred
You will still need to soak the tissue before cutting, but it gives good
infiltration and soaking times are decreased.
James Watson HT, ASCP
Facilities Manager of Histology
GNF, Genomics Institute of the Novartis Research Foundation
Room C015
858-332-4647
jwatson <@t> gnf.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gladney,
Diane C Ms MACH
Sent: Monday, February 12, 2007 3:57 AM
To: Pam V; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Softening animal tissue
Pam,
What is the dilution of glycerin (strength) that you use? I would be
interested in trying this out. Any other guidance such as how long do
you soak your blocks, etc would be helpful also.
Thanks,
Diane
Diane C. Gladney, HT (ASCP)
Supervisor, Anatomical Pathology
Moncrief Army Community Hospital
Dept. of Pathology
P.O. Box 484
4500 Stuart St.
Ft. Jackson, SC 29207
Email: diane.gladney <@t> se.amedd.army.mil
Phone: 803-751-2530
FAX: 803-751-7829
DSN: 734-2530
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Pam V
Sent: Sunday, February 11, 2007 7:46 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Softening animal tissue
Hi all...
One of the things I've done for years is soak tissue with a dilute
glycerin. Not exactly sure why, but the cells do not blow up, nor is the
staining affected, neither routine nor immunos..Sections adhere to the
slides just fine.
I'm at Evanston Northwestern Hospital in Evanston Illinois...we do some
work on animal tissue also and this has worked well for me. It also
works on decals, and very well on blood, such as bone marrow clots, as
well as lymph nodes. It also allows thinner sectioning.
I haven't done any recent searches for the use of glycerin and some
people are hesitant, but I did see that it's suggested by Frieda Carson
and is a question in the ASCP Board of Registry practice exam text.
Pam Vlies HTASCP
ENH Histology lab..
----- Original Message ----
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<histonet-request <@t> lists.utsouthwestern.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Sunday, February 11, 2007 11:59:26 AM
Subject: Histonet Digest, Vol 39, Issue 18
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Today's Topics:
1. Re: Animal Tissue (Esther Peters)
2. Re: Animal Tissue (Amos Brooks)
3. Re: Iron Control (Jennifer MacDonald)
4. Job opening (raj)
5. GPR58 antibody (Ana MERINO-TRIGO)
----------------------------------------------------------------------
Message: 1
Date: Sat, 10 Feb 2007 14:43:28 -0500
From: Esther Peters <esther.peters <@t> verizon.net>
Subject: Re: [Histonet] Animal Tissue
To: jhnspam <@t> aol.com
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <45CE2060.70203 <@t> verizon.net>
Content-Type: text/plain; charset=us-ascii; format=flowed
Pam,
What animal tissue are you using? I recall learning from HistoNet a
while back that rodent tissues should be processed for shorter periods
(30 min. per solution and paraffin change compared to 60 min.) because
of potential hardening issues. This is also noted in several mouse/rat
or small animal processing schedules provided in the Animal Processing
Manual published by the National Society for Histotechnology's
Veterinary, Industry and Research Committee, edited by Gayle Callis and
Diane Sterchi (along with many other helpful tips for non-human tissue
handling!). I think it (or an updated version) is still available from
NSH (Gayle, Diane?).
Esther Peters, Ph.D.
George Mason University
jhnspam <@t> aol.com wrote:
> I need to get some input on cutting animal tissue. I have always
worked in a
> clinical setting and have recently moved into a research lab. I find
that the
> tissue is very brittle and needs to be iced for an extremely amount
of time.
> Can any of you animal cutting histo techs please give me some advice
on what
> type of paraffin you are using and what processing schedule you are
using. I
> have recently changed to Richard Allen type 9 paraffin and it seems
to have
> helped some. I would appreciate any advice you can give me.
>
> Thanks,
> Pam
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
------------------------------
Message: 2
Date: Sat, 10 Feb 2007 17:22:48 -0500
From: "Amos Brooks" <amosbrooks <@t> gmail.com>
Subject: Re: [Histonet] Animal Tissue
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<582736990702101422u355b9e06rd0c40e0c575e8e97 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Hi,
If anyone has any replies to this please do it to the list (or
include me as well) as I seem to be in a similar boat having recently
accepted a transfer (within the same company) to a research lab. I must
admit to some trepidation about animal tissue not having much experience
with it. My experiences are primarily clinical. I've heard some stories
of great frustration about animal tx. I'm sure I can handle it but there
is always a learning curve.
Thanks
Amos Brooks
Message: 19
Date: Fri, 9 Feb 2007 22:57:36 EST
From: jhnspam <@t> aol.com
Subject: [Histonet] Animal Tissue
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <c65.c5cc97a.32fe9cb0 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"
I need to get some input on cutting animal tissue. I have always worked
in a clinical setting and have recently moved into a research lab. I
find that the tissue is very brittle and needs to be iced for an
extremely amount of time. Can any of you animal cutting histo techs
please give me some advice on what type of paraffin you are using and
what processing schedule you are using. I have recently changed to
Richard Allen type 9 paraffin and it seems to have helped some. I would
appreciate any advice you can give me.
Thanks,
Pam
------------------------------
Message: 3
Date: Sat, 10 Feb 2007 17:49:46 -0800
From: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>
Subject: Re: [Histonet] Iron Control
To: "sheila adey" <sheila_adey <@t> hotmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF18F6358F.443EAEEB-ON8825727F.000A0CF8-8825727F.000A0CFA <@t> mtsac.edu>
Content-Type: text/plain; charset="UTF-8"
Spleen makes for a sensitive iron control. The amount of ir
sufficient, but not overwhelming.
Jennifer
-----histonet-bounces <@t> lists.utsou
To: histonet <@t> lists.utsouthwestern.edu
From: "sheila Sent by: histonet-bounces <@t> lists. Date:
02/10/2007 07:18AM
Subject: [Histonet] Iron Hello All,
We are almost out of Iron Control. We h chronic
hemosiderosis condition. The Docs sa before we
find a suitable control. Could any that might
be easier to find?
Th Sheila Adey HT MLT
Port Huron Hospital
Mic ____________________ ______________________ 5F__
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Your Space. Windows Live
Spaces. _____ ______________________ 5F__ Histonet
mailing lis Histonet <@t> lists.utsouthwestern.edu
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References
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------------------------------
Message: 4
Date: Sat, 10 Feb 2007 21:58:51 -0500
From: raj <raj <@t> bluemarble.net>
Subject: [Histonet] Job opening
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <45CE866B.3B47CBA9 <@t> bluemarble.net>
Content-Type: text/plain; charset=us-ascii
We have a opening at Bloomington Hospital for a reg. HT. Bloomington,
IN. It is a day shift job. Bloomington is about 50 miles south of
Indianapolis,IN. It the home of Indiana University. If interested
please reply and I will direct you to the HR dept. Thank You Rebecca A.
Johnson
------------------------------
Message: 5
Date: Sun, 11 Feb 2007 17:09:40 +0100 (CET)
From: Ana MERINO-TRIGO <ana.merino-trigo <@t> wanadoo.fr>
Subject: [Histonet] GPR58 antibody
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <24357752.403521171210180349.JavaMail.www <@t> wwinf1603>
Content-Type: text/plain; charset=UTF-8
Hello Histonet,
I was wondering if anyone has experience with GPR58, G protein coupled
receptor (alias TAAR2, trace amine associated receptor 2) human antibody
from LifeSpan using Ventana. So far, I've had no luck on paraffin
sections. I will need to test the expression on human pancreas (paraffin
sections). I'm using human cerebelum as positive control, as it's
described in the literature to be expressed in this tissue.
I've tested, CC1, CC2 and protease using DAB kit for the developing.
With CC2 I do lose mostly of my tissues, no sure if is a buffer batch
problem or if conditions are really strong for cerebellum tissue.
Any advice it will be really appreciate it.
thanks a lot,
Ana
------------------------------
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