[Histonet] Freezing human skin biopsies for cytokine ICC
Downs, Heather M.
HDOWNS <@t> PARTNERS.ORG
Fri Feb 16 12:08:38 CST 2007
We often section human skin punch biopsy, using a sliding microtome and dry ice.
We fix our tissue for 12 hours in PLP, followed by a wash in Sorenson's and
placed in croprotectant for 24 hours before either cutting it, or freezing @-20.
We do a free floating immunostain and cut at 50 microns with no problems.
HD
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Sent: Friday, February 16, 2007 1:01 PM
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Subject: Histonet Digest, Vol 39, Issue 26
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Today's Topics:
1. RE: Thermometers (Liz Chlipala)
2. Looking for Contact (Steven P Postl)
3. primary incubation times (MKing)
4. New Jersey Symposium (Joseph Tamasi)
5. RE: Tissue Microarray tools (Annette Hall)
6. Jacquelyn Grewe/Staff/OhioHealth is out of the office .
(JGREWE <@t> OhioHealth.com)
7. Freezing human skin biopsies for cytokine ICC (Jamie E Erickson)
8. gelatinized slides (Atoska Gentry)
9. HT position in Seattle (ckduffynw <@t> aol.com)
10. Re: Thermometers (soofia siddiqui)
----------------------------------------------------------------------
Message: 1
Date: Thu, 15 Feb 2007 11:16:54 -0700
From: "Liz Chlipala" <liz <@t> premierlab.com>
Subject: RE: [Histonet] Thermometers
To: "'Lester Raff'" <LRaff <@t> lab.uropartners.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001c7512d$7af456c0$0d00a8c0 <@t> domain.Premier>
Content-Type: text/plain; charset="us-ascii"
It depends upon your SOP's most calibrated thermometers have a little
statement at the bottom of the certification that states that most
institutions recalibrate once a year but you must follows your own SOP's.
You then need to take into consideration what guidelines you are following
and what the standards are. What I'm saying is that you can write in your
SOP's that you are going to calibrate once every 5 years, but when you get
inspected will the inspector think that is good enough. The kicker is the
cost to recalibrate is more than a new thermometer, we just purchase a new
calibrated thermometer each year and that's what is in our SOP's.
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
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lester Raff
Sent: Thursday, February 15, 2007 10:31 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Thermometers
Do labs generally recalibrate/recertify NIST calibrated thermometers?
If so, how and how often?
Lester J. Raff, MD
Medical Director
UroPartners, LLC Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, IL 60154
ph: 708-486-0076
fax: 708-486-0080
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Message: 2
Date: Thu, 15 Feb 2007 12:58:43 -0600
From: Steven P Postl <steven.p.postl <@t> abbott.com>
Subject: [Histonet] Looking for Contact
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF918D180C.9B59A05C-ON86257283.00682301-86257283.00684B62 <@t> abbott.com>
Content-Type: text/plain; charset="US-ASCII"
If Mary Gessford is out there, could you please contact me? Thanks.
------------------------------
Message: 3
Date: Thu, 15 Feb 2007 14:17:16 -0500
From: MKing <making <@t> ufl.edu>
Subject: [Histonet] primary incubation times
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <45D4B1BC.2050305 <@t> ufl.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Patrick,
I remember reading in one of the early immunohistochemistry texts that
the reaction between a primary antibody and its epitope was believed to
require 96 hrs. at 4 deg. C to reach equilibration. Whether or not
that's true for your antibodies, it is the kind of precedent that
underlies the procedures typically used today.
The other issue is penetration--if you need tissue to be evenly labeled
throughout the depth of the specimen then section thickness and
embedding material may require long incubations (at least). If surface
labeling of an abundant epitope is enough to tell you what you want to
know, a good primary may require less than half an hour!
Happy histo,
Mike King
UF Neuroscience
----------------------------------------
Date: Wed, 14 Feb 2007 15:24:53 -0800
From: "Patrick Laurie" <plaurie <@t> benaroyaresearch.org>
Subject: [Histonet] Reason for lengthy primary antibody incubation?
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <420f1b520f9ee44994edfc63ad644f47 <@t> localhost.localdomain>
Content-Type: text/plain; charset="us-ascii"
------------------------------
Message: 4
Date: Thu, 15 Feb 2007 15:26:10 -0500
From: Joseph Tamasi <joseph.tamasi <@t> bms.com>
Subject: [Histonet] New Jersey Symposium
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <45D4C1E2.208 <@t> bms.com>
Content-Type: text/plain; charset="iso-8859-1"
Dear Histonet Subscribers that live in or around New Jersey:
The New Jersey Society for Histotechnology is offering a two day
symposium on March 16-17, 2007 at the Clarion Hotel and Conference
Center in Cherry Hill, NJ. You can view the complete program and get
registration information on the NSH website. Go to www.NSH.org, then on
the left hand side of the page click on NSH Regions / Meeting Calendar /
Region II / NJSH Symposium. Scroll down for the link to view the full
program.
------------------------------
Message: 5
Date: Thu, 15 Feb 2007 14:50:13 -0600
From: Annette Hall <Annette_hall <@t> pa-ucl.com>
Subject: RE: [Histonet] Tissue Microarray tools
To: 'Akemi Allison-Tacha' <akemiat3377 <@t> yahoo.com>, Helen Fedor
<hfedor <@t> jhmi.edu>
Cc: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<9FC023A4AB52BB4D87DC6456081A822C012B5AD8 <@t> mercury.pa-ucl.com>
Content-Type: text/plain; charset="iso-8859-1"
I just received the catalog yesterday and was thrilled to see this product.
Although a bit pricey-$1530, we should be able to capture the cost with
reagent and materials savings, as well as tech time.
Thanks to all of you who responded to my inquiry,
Annette
-----Original Message-----
From: Akemi Allison-Tacha [mailto:akemiat3377 <@t> yahoo.com]
Sent: Thursday, February 15, 2007 9:58 AM
To: Helen Fedor
Cc: histonet
Subject: Re: [Histonet] Tissue Microarray tools
Helen,
I just saw a new "TMA Builder" that Lab Vision has available cat# TMA-001
under Research Tools in their new catalog. It looks similar to what
woorie-medic's tissue punch extractor looked like. The mold is totally
different though, it has a metal clamp & a metal recipient base mold with 24
core indentations. Have you seen it? Any comments?
Akemi Allison-Tacha BS, HT(ASCP)HTL
President
Phoenix Lab Consulting & Staffing
Specializing in Histology, IHC & TMA
Madison, WI
Cell: (925)788-0900
E-mail: akemiat3377 <@t> yahoo.com
Helen Fedor <hfedor <@t> jhmi.edu> wrote:
Dear Annette, We have been using the Beecher system and have made over
450 TMA's with the instrument, It is reliable and durable.
Best Regards,
Helen
Helen L. Fedor B.S.
Johns Hopkins University
Pathology Department
600 N Wolfe St
Marburg Room 406
Baltimore MD 21287
email: hfedor <@t> jhmi.edu
Phone: 410 614-1660
Pager: 410 283-3419
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>>> Annette Hall 2/13/2007 2:32 PM >>>
Can anyone recommend a good system for making tissue microarrays. We
are not
a large volume lab (approx 250 IHC per month), so I would be looking
at
manual methods for accomplishing this. My planned uses would be for
new
antibody evaluations and controls.
Thanks, Annette
Annette J Hall, MT
Micro/Cyto/Histo Supervisor
United Clinical Labs
205 Bluff Street
Dubuque, IA 52001
Phone: 563.556.2010
Cell: 563.580.9751
Fax: 563.584.2085
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------------------------------
Message: 6
Date: Thu, 15 Feb 2007 16:02:50 -0500
From: JGREWE <@t> OhioHealth.com
Subject: [Histonet] Jacquelyn Grewe/Staff/OhioHealth is out of the
office .
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF2D255089.BA3DA97D-ON85257283.00739DF7-85257283.00739DF7 <@t> ohiohealth.com>
Content-Type: text/plain; charset=US-ASCII
I will be out of the office starting 02/14/2007 and will not return until
02/19/2007.
I will respond to your message when I return. Thanks, Jackie
------------------------------
Message: 7
Date: Thu, 15 Feb 2007 17:40:24 -0500
From: Jamie E Erickson <jamie.erickson <@t> abbott.com>
Subject: [Histonet] Freezing human skin biopsies for cytokine ICC
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OFA75B1E3B.87205A07-ON85257283.007BAA7B-85257283.007C8C8B <@t> abbott.com>
Content-Type: text/plain; charset="US-ASCII"
Hello histonetters,
I hope someone can help me with a
question about how to freeze a human skin biopsy. I am in research but am
in collaboration with a clinical lab that will be collecting human skin
biopsies for me to do cytokine ICC of TNF,IFN-g, among others and
possibly cell makers.
My question is how do I instruct them in freezing the skin so that
I get the best sample for this staining. I am going to embed the sample
once it is in my lab for cryosectioning. Also if people know of
manufactures of antibodies that sell TNF,IFN-g that work in human tissue
that would also be helpful, I have found a few but many different
protocols and fixations.
Any thoughts how be helpful.
Jamie
_______________________________
Jamie Erickson
Sr. Research Associate
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com
------------------------------
Message: 8
Date: Thu, 15 Feb 2007 17:01:45 -0600
From: Atoska Gentry <gentras <@t> vetmed.auburn.edu>
Subject: [Histonet] gelatinized slides
To: Histonet <histonet <@t> pathology.swmed.edu>
Message-ID: <45D4E659.1070909 <@t> vetmed.auburn.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
hello, one of our collaborating researchers has me searching for
gelatinized slides to be used for Golgi staining for cryosectioning 100
micron mouse
spines. It appears that she came up with this idea after reading about
a similar protocol on brain. I have searched the histonet archives and
the only thing that I can come up with is info on gelatin subbed slides.
Thus far I've only found one company that has commercial gelatinized
slides. Please pardon my ignorance but is there a significant difference
between the two whereas one is preferable for certain protocols for
which the other is not? And if any of you are currently using commercial
gelatinized slides will you provide me with contact info on your source?
Thanks, Atoska
--
Atoska S. Gentry, B.S., HT(ASCP)
Research Assistant IV
Scott-Ritchey RSCH Center
College of Vet. Med
Auburn, AL 36849
PH (334) 844-5579
FAX (334) 844-5850
email: gentras <@t> vetmed.auburn.edu
------------------------------
Message: 9
Date: Fri, 16 Feb 2007 11:34:17 -0500
From: ckduffynw <@t> aol.com
Subject: [Histonet] HT position in Seattle
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <8C91FFF6FD678A6-764-19B2 <@t> WEBMAIL-RC06.sysops.aol.com>
Content-Type: text/plain; charset="us-ascii"
One part-time and one full time position needed immediately in Edmonds area.
ASCP cert required. Small lab, excellent benefits, competitve salary. Contact
Kay at ckduffynw <@t> aol.com.
________________________________________________________________________
Check out the new AOL. Most comprehensive set of free safety and security
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------------------------------
Message: 10
Date: Fri, 16 Feb 2007 09:00:17 -0800 (PST)
From: soofia siddiqui <soofias2 <@t> yahoo.com>
Subject: Re: [Histonet] Thermometers
To: Lester Raff <LRaff <@t> lab.uropartners.com>,
histonet <@t> lists.utsouthwestern.edu
Message-ID: <201359.87917.qm <@t> web39507.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Initially I found to get new thermometer each year was easier and economical,
but after some research I came up with a cheaper solution. Now in our lab we
recalibrate NIST calibrated thermometer every year. We keep two sets of
thermometers for each freezer and fridge and recalibrate and use them alternate
year.One which is currently in use is just got calibrated in January,07and will
expire in January,08. The other one which is expiring in Feb ,07 we will send
for recalibration in December ,07 which will come back by January,08 and we will
use recalibrated one for the next year. We send our thermometer.to the following
company. . Calibration Services
Control Company
4455 Rex Road
Friendswood, TX 77546
They charge $15 for each thermometer and $8 for shipping. We get two
thermometer done in $50 including shipping from both ways. I found it cheaper
that way then buying new thermometer each year. I believe that calibration can
be done in house too. I am sure there are very smart people out there in this
field who may have better ways to deal with this issue.
Soofia
Lester Raff <LRaff <@t> lab.uropartners.com> wrote:
Do labs generally recalibrate/recertify NIST calibrated thermometers?
If so, how and how often?
Lester J. Raff, MD
Medical Director
UroPartners, LLC Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, IL 60154
ph: 708-486-0076
fax: 708-486-0080
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