[Histonet] RE: Histonet Ventana staining Avidin Biotin block
White, Lori
lwhite <@t> lakeridgehealth.on.ca
Tue Mar 23 14:42:10 CST 2004
Hi,
We use the avidin/block on the Ventana Nexes as necessary and it works very
well. I don't know that it would be advantageous to use for all tissues,
considering the extra time that would be built into the run.....
Lori
-----Original Message-----
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Sent: Tuesday, March 23, 2004 1:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 4, Issue 27
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Today's Topics:
1. Atoska Gentry/ B Plus (Joyce Cline)
2. Ventana staining Avidin Biotin Block (Featherstone, Annette)
3. Re: Ventana staining Avidin Biotin Block (Lilia Muolo)
4. RE: Ventana staining Avidin Biotin Block (GUTIERREZ, JUAN)
5. Wendy England/ iron smears (Joyce Cline)
6. IHC position in San Francisco bay area. (Morken, Tim - Labvision)
7. Small Animal atlas (Volumes 1 and 2) are back in print!
(Gayle Callis)
8. RE: Phosphotungstic Acid Hematoxylin stain without usi ng
Zenker's (Tony Henwood)
9. please remove me from your mailing list (Anna Bozzano)
10. RE: Web site for Cytology (Kemlo Rogerson)
11. subscribe (Anne Van Binsbergen)
12. Mouse Lung (Starkus, Laurie)
13. RE: help with elastic staining (Featherstone, Annette)
14. Problems with c-fos immediate early gene labelling (komo <@t> bu.edu)
15. RE: CD98 and PLGF (Featherstone, Annette)
16. Re: Problems with c-fos immediate early gene labelling
(Geoff McAuliffe)
17. FW: Job opening - Wildlife Conservation Society (McAloose, Dee)
18. IHC on blood and cytology smears (ICC) (Jan Shivers)
19. NYSHS 2004 Annual Meeting (Luis Chiriboga)
----------------------------------------------------------------------
Message: 1
Date: Mon, 22 Mar 2004 13:15:40 -0500
From: "Joyce Cline" <jcline <@t> wchsys.org>
Subject: [Histonet] Atoska Gentry/ B Plus
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <006901c41039$af661660$1d2a14ac <@t> wchsys.org>
Content-Type: text/plain; charset="iso-8859-1"
Hi Atoska
I get my B Plus from Market Lab, order # ML0407, Phone # 800-237-3604.
I do not know it's components, other than formaldehyde. The company that
makes it is Advanced Biomedical Reagents & Technologies.
------------------------------
Message: 2
Date: Mon, 22 Mar 2004 13:47:13 -0500
From: "Featherstone, Annette" <AFeatherstone <@t> KaleidaHealth.Org>
Subject: [Histonet] Ventana staining Avidin Biotin Block
To: "Histonet (E-mail)" <histonet <@t> pathology.swmed.edu>
Message-ID:
<DF6AB9298498D31199CF0008C7E6C1200CF95C53 <@t> kalmb02.kaleidahealth.org>
Content-Type: text/plain; charset="iso-8859-1"
hi
Just wondering if anyone out there using the Ventana can tell me if they are
using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system
antibodies. Our institution is only using them on a select few and I think
we should use it on all of them.Thanks for your response
Annette Featherstone HT/MLT
Kaleida Health
Buffalo General Hospital
100 High St
Buffalo NY 14203
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------------------------------
Message: 3
Date: Mon, 22 Mar 2004 14:21:12 -0500
From: Lilia Muolo <muololi <@t> umdnj.edu>
Subject: Re: [Histonet] Ventana staining Avidin Biotin Block
To: AFeatherstone <@t> KaleidaHealth.Org, histonet <@t> pathology.swmed.edu
Message-ID: <s05ef679.070 <@t> smtpnpc.umdnj.edu>
Content-Type: text/plain; charset=US-ASCII
Hi, our lab only uses the A /B blocking on the Ventana if necessary but
usually we don't need to use it. We use the Discovery here but I do
remember using the A/B blocking more often when we had the Nexes. I
would imagine that the Benchmark would be like the Discovery.
Lil Muolo
Cancer Institute of New Jersey
>>> "Featherstone, Annette" <AFeatherstone <@t> KaleidaHealth.Org> 3/22/04
1:47:13 PM >>>
hi
Just wondering if anyone out there using the Ventana can tell me if
they are
using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC
system
antibodies. Our institution is only using them on a select few and I
think
we should use it on all of them.Thanks for your response
Annette Featherstone HT/MLT
Kaleida Health
Buffalo General Hospital
100 High St
Buffalo NY 14203
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------------------------------
Message: 4
Date: Mon, 22 Mar 2004 13:21:59 -0600
From: "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org>
Subject: RE: [Histonet] Ventana staining Avidin Biotin Block
To: "Featherstone, Annette" <AFeatherstone <@t> KaleidaHealth.Org>,
"Histonet (E-mail)" <histonet <@t> pathology.swmed.edu>
Message-ID:
<A61F23E937E4DA488E0C0F60093843D90F6A2B <@t> ccetxm030.echristus.net>
Content-Type: text/plain; charset="utf-8"
We use it on everything. It's a lot easier than having a different protocol
for aech type of tissue.
Juan
-----Original Message-----
From: Featherstone, Annette [mailto:AFeatherstone <@t> KaleidaHealth.Org]
Sent: Mon 3/22/2004 12:47 PM
To: Histonet (E-mail)
Cc:
Subject: [Histonet] Ventana staining Avidin Biotin Block
hi
Just wondering if anyone out there using the Ventana can tell me if
they are
using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC
system
antibodies. Our institution is only using them on a select few and
I think
we should use it on all of them.Thanks for your response
Annette Featherstone HT/MLT
Kaleida Health
Buffalo General Hospital
100 High St
Buffalo NY 14203
CONFIDENTIALITY NOTICE:
This email transmission and any documents, files,
or previous e-mail messages attached to it are
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ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777.
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------------------------------
Message: 5
Date: Mon, 22 Mar 2004 15:56:30 -0500
From: "Joyce Cline" <jcline <@t> wchsys.org>
Subject: [Histonet] Wendy England/ iron smears
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001f01c41050$277dcb00$1d2a14ac <@t> wchsys.org>
Content-Type: text/plain; charset="iso-8859-1"
I use an old fashioned way of fixing the bone marrow smears. We had trouble
with no reaction on our smears. Now we use a coplin jar with a small piece
of paper towel in the bottom, drop 10% buffered formalin to saturate the
paper towel but not to reach the blood on the slides. Leave the slides in
for 10 minutes and stain normally. We have not had any trouble with an iron
reaction since using this method. But do not leave the slides in more than
15 minutes max.
------------------------------
Message: 6
Date: Mon, 22 Mar 2004 14:30:56 -0800
From: "Morken, Tim - Labvision" <tpmorken <@t> labvision.com>
Subject: [Histonet] IHC position in San Francisco bay area.
To: "Histology Net List Server (E-Mail)"
<histonet <@t> pathology.swmed.edu>
Message-ID:
<0556BE8AC5551E4E8AF6BB9E42509BA217741F <@t> usca0082k08.labvision.apogent.com>
Content-Type: text/plain
Lab Vision, a fast-growing biotech company located in Fremont, California,
just southeast of San Francisco, has an opening for an Immunohistochemistry
Technologist in the Quality Control laboratory. The position entails testing
of manufactured lots of antibody and detection reagents as well as
prospective reagents. The applicant must have extensive experience in
immunohistochemistry, especially in development of new antibodies and other
IHC reagents. Advancement is limited only by personal initiative.
Certification by ASCP as HT or HTL and/or QIHC is preferred. Salary is
negotiable.
Lab Vision (www.labvision.com) is a world-leading manufacturer of
IHC-automation instruments and immunohistochemistry reagents for the
pathology laboratory and biological research. Lab Vision is an Apogent
company (www.apogent.com)
If interested please send resume by email to:
Tim Morken
Product Development
Lab Vision / Neomarkers
47790 Westinghouse Dr.
Fremont, CA 94539
USA
PH: 510-991-2840
FAX: 510-991-2826
email: tpmorken <@t> labvision.com
www.labvision.com
------------------------------
Message: 7
Date: Mon, 22 Mar 2004 16:12:28 -0700
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: [Histonet] Small Animal atlas (Volumes 1 and 2) are back in
print!
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <3.0.6.32.20040322161228.00bcc478 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"
Dear All,
For those of you who work with or dissect laboratory animals and are in
need of an atlas to guide the way.
After many years of frustration and sadness when these atlases went out of
print, they are back on the market. I saw the new atlas today, and couldn't
wait to order it. A huge silent hooray!
There are two color atlases (volume 1 is for rabbit and guinea pig and
Volume 2 is rat, mouse, and Golden hamster. Pricing for Vol 1 and Vol 2
are $139 per atlas, and that is a deal.
ISBN:0702027030
Title:Colour Atlas of Anatomy of Small Laboratory Animals, Volume 2, mouse,
rat, hamster
Price:$139.00
Go to Elsevier website to order, it is very user friendly, excellent.
I just gave/ordered myself an early birthday gift!
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 8
Date: Tue, 23 Mar 2004 11:22:18 +1100
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] Phosphotungstic Acid Hematoxylin stain without
usi ng Zenker's
To: "'Boswell'" <kcboswell <@t> grandecom.net>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E16A <@t> simba.kids>
Content-Type: text/plain; charset="iso-8859-1"
Dear Boswell,
The following Cherukian's modification works very well.
Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318
http://www.histosearch.com/homepages/TonyHenwood/default.html
http://us.geocities.com/tonyhenwoodau/index.html
Document Procedure:
Phosphotungstic Acid Haematoxylin
Principle:
Cherukian's modification employs an eosin solution that stains the
erythrocytes red and differentiates them from the blue fibrin.
Fixation: 10% buffered formalin.
Microtomy: paraffin sections at 5um.
Controls:
Use brain sections and section of muscle. A good stain will demonstrate the
dendrites as blue where as in a bad stain they appear light grey to salmon
in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and
collagen appear red. Muscle striations should be well defined.
Reagents:
1. Eosin: - Warning: Flammable - see MSDS
Eosin Y, water soluble (CI 145380) 0.5g
Distilled Water 10ml
80% Ethanol 190ml
Working: 10ml stock and
Before use add 50ul glacial acetic acid.
2. 1% Periodic Acid
3. PTAH solution
Haematoxylin (CI 75290) 0.5g
Phosphotungstic Acid 10g
Distilled water 500ml
Dissolve solid ingredients in separate portions of the water. Use gentle
heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium
permanganate to ripen. The stain is ready to use.
Procedure:
1. Dewax and hydrate sections to 80% alcohol.
2. Place slides in eosin for 30 seconds.
3. Wash slides in distilled water for a few seconds.
4. Place slides in 1% periodic acid for 20 minutes.
5. Wash slides in water for 3 minutes.
6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30
minutes on.
7. Dehydrate, clear and mount.
Results:
Dendrites, nuclei, fibrin, platelets and muscle - blue
Red blood cells and collagen - red.
Notes:
Reference:
1. Cherukian, C.J., Histologic. 8(4); 105, (1977).
2. Luna, L., Histologic. 5(2); 66, (1975).
-----Original Message-----
From: Boswell [mailto:kcboswell <@t> grandecom.net]
Sent: Saturday, 20 March 2004 7:30 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using
Zenker's
Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain
without using Zenker's. We have looked everywhere and are unable to find
it. Thank you.
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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------------------------------
Message: 9
Date: Tue, 23 Mar 2004 10:14:40 +0100
From: Anna Bozzano <bozz <@t> icm.csic.es>
Subject: [Histonet] please remove me from your mailing list
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <3.0.1.32.20040323101440.0120fd88 <@t> cucafera.icm.csic.es>
Content-Type: text/plain; charset="us-ascii"
please remove me from your mailing list
------------------------------
Message: 10
Date: Tue, 23 Mar 2004 09:21:37 -0000
From: "Kemlo Rogerson" <kemlo <@t> tiscali.co.uk>
Subject: RE: [Histonet] Web site for Cytology
To: "'Kathleen Boozer'" <BoozerKA <@t> pa1.ah.org>,
<histonet <@t> pathology.swmed.edu>
Message-ID: <000001c410b8$3f376b80$48362850 <@t> KEMLOS>
Content-Type: text/plain; charset="us-ascii"
Cytopathnet.org I think. It sort of comes and goes depending on the
variety of virus it becomes infected with.
I'm always amused that it becomes regularly infected by viruses; sort of
gives you a feeling of its authenticity. I think it is negative for the
oncogenic variety but hope it has regular tests including a silicone
biopsy.
Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS
Tel: 0208 970 8414
Mob: 07830 196072
Mobile E-Mail kemlorogerson <@t> 3mail.com
FAX & Answer Phone 0871 242 8094
E-mail Accounts:
kemlo <@t> tiscali.co.uk or
kemlo1 <@t> btinternet.com
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kathleen
Boozer
Sent: 17 March 2004 17:13
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] Web site for Cytology
Is there something like the Histonet available for Cytology? I have a
co-worker looking for perameters in quality control for her department
as she documents screening errors. Thanks!
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 11
Date: Tue, 23 Mar 2004 12:59:07 +0400
From: "Anne Van Binsbergen" <vanann702 <@t> skmc.gov.ae>
Subject: [Histonet] subscribe
To: <histonet <@t> pathology.swmed.edu>
Message-ID:
<0C44F1AAEE47D54DA4210A60AB206F5E01F18A95 <@t> SKMCEMAIL.skmc.gov.ae>
Content-Type: text/plain; charset="iso-8859-1"
------------------------------
Message: 12
Date: Tue, 23 Mar 2004 07:56:38 -0500
From: "Starkus, Laurie" <StarkusL <@t> ummhc.org>
Subject: [Histonet] Mouse Lung
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<E9F72FA12FC6D411AD750006298F2F8205878A0F <@t> hcunivexch04.ummhc.org>
Content-Type: text/plain; charset="ISO-8859-1"
I've recently started working with mouse lung and was given a protocol for
processing. The protocol calls for 0.85% NaCl for 60 minutes at 4 degrees
celsius. Then a 1:1 0.85% NaCl to Absolute ethanol for 30 minutes at room
temp. Then the processing looks rather normal. Since my experience is with
human tissue, I've never seen a sodium chloride step in processing. What
does it do? And, is it really necessary? This is mouse lung infected with
TB or cryptococcus.
Thanks in advance.
------------------------------
Message: 13
Date: Tue, 23 Mar 2004 08:34:14 -0500
From: "Featherstone, Annette" <AFeatherstone <@t> KaleidaHealth.Org>
Subject: RE: [Histonet] help with elastic staining
To: 'Patsy Ruegg' <pruegg <@t> colobio.com>, Deb Stults
<DSTULTS <@t> fremontmemorial.org>, histonet <@t> lists.utsouthwestern.edu
Message-ID:
<DF6AB9298498D31199CF0008C7E6C1200CF95C54 <@t> kalmb02.kaleidahealth.org>
Content-Type: text/plain; charset="iso-8859-1"
When doing the pentachrome stain do not over differentiate the elastic too
much in the ferric chloride, keep it kind of dark. The acid solutions that
follow continue to lighten the elastic fibers.
Annette FeatherstoneHT/MLT
Kaleida Health
-----Original Message-----
From: Patsy Ruegg [mailto:pruegg <@t> colobio.com]
Sent: Friday, March 19, 2004 14:19
To: Deb Stults; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] help with elastic staining
I have this problem when I do a Pentachrome stain for elastic fibers, the
fibers stain fine when I first do the hematoxylin but as I continue with the
rest of the stain the fiber stain is lost, I have remedied this by repeating
the hematoxylin again at the end of the staining process to get back the
fiber staining. Try it.
Patsy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Deb
Stults
Sent: Friday, March 19, 2004 11:48 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] help with elastic staining
We ran into a problem today with an elastic stain on an artery biopsy.
We're pretty sure it is due to a reagent problem, but not sure which one.
The elastic fibers did not stain at all. We use Weigert's iron hematoxylin
solutions A and B, which are new bottles and worked yesterday in another
stain (we make up fresh before each use), the Resorcin Fuchsin working
solution expired last week, and our new bottle (previously ordered) has not
yet arrived, and the Van Gieson's solution is brand new just opened today.
Since the fibers are not black, that makes me think that its the Weigert's,
but since the Resorcin Fuchsin is expired, we weren't sure if it could be
the problem. If anyone has any suggestions, please let me know asap. The
pathologist doesn't want to wait for new reagent to come in.
Thanks,
Deb and Karen
_______________________________________________
Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
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CONFIDENTIALITY NOTICE:
This email transmission and any documents, files,
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responsible for delivering it to the intended recipient,
you are hereby notified that any further review,
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If you have received this message in error, please
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ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777.
------------------------------
Message: 14
Date: Tue, 23 Mar 2004 09:48:31 -0500
From: komo <@t> bu.edu
Subject: [Histonet] Problems with c-fos immediate early gene labelling
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <1080053311.40604e3f585a5 <@t> www.bu.edu>
Content-Type: text/plain
I'm hoping I could get some helpful advice on what might cause my c-fos
IHC labelling to work well in one lab, but not my own. I am running the
protocol on rat brains using a hydrogen peroxide/methanol step for
endogenous peroxidase blocking, a normal goat serum blocking step, a
c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated
secondary, a streptavidin horeradish peroxidase, and a nickle enhanced
DAB reaction.
Unfortunately, although I have all of the exact same antibodies, chemicals,
and concentrations as the lab from which the protocol was developed, the
label is weak and not very specific when I run it at my home lab. In fact, I
even took brain slices from the same rats that I was running at my lab and
ran them at the lab where the protocol was developed, and found that the
c-fos label was beautiful with a low background.
Does anyone has any suggestions as to why this might be? One person
at the other lab suggested that possibly the wells that we run the tissue in
were not being cleaned properly (we soak them in bleach while the other
lab uses just pex). Could this have any effect?
Thanks for any advice you can provide!!
Rob Komorowski
------------------------------
Message: 15
Date: Tue, 23 Mar 2004 09:50:21 -0500
From: "Featherstone, Annette" <AFeatherstone <@t> KaleidaHealth.Org>
Subject: RE: [Histonet] CD98 and PLGF
To: 'Patsy Ruegg' <pruegg <@t> colobio.com>, "Histonet <@t> Pathology. Swmed.
Edu" <histonet <@t> pathology.swmed.edu>
Cc: "Ihcrg <@t> Yahoogroups. Com" <ihcrg <@t> yahoogroups.com>
Message-ID:
<DF6AB9298498D31199CF0008C7E6C1200CF95C55 <@t> kalmb02.kaleidahealth.org>
Content-Type: text/plain; charset="iso-8859-1"
YES!!I am! Or was...
I used a pepsin pretreatment @37* for 15 minutes, Block with a protein
block, primary antibody 1:100 for 1hr, Goat secondary 30 min rt, Alk phos 30
minutes rt, and whatever chromgen you use for alk phos, rt 40 min. My Plgf
is from Santa Cruz.
Annette Featherstone HT/MLT
-----Original Message-----
From: Patsy Ruegg [mailto:pruegg <@t> colobio.com]
Sent: Friday, March 19, 2004 13:58
To: Patsy Ruegg; Histonet <@t> Pathology. Swmed. Edu
Cc: Ihcrg <@t> Yahoogroups. Com
Subject: RE: [Histonet] CD98 and PLGF
I guess no one is using cd98 and/or Placental Growth Factor on ffpe
tissue????
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Patsy
Ruegg
Sent: Wednesday, March 17, 2004 10:03 AM
To: Histonet <@t> Pathology. Swmed. Edu
Cc: Ihcrg <@t> Yahoogroups. Com
Subject: [Histonet] CD98 and PLGF
I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160
and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies
work in ffpe tissue?
Patsy
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Message: 16
Date: Tue, 23 Mar 2004 10:24:38 -0800
From: Geoff McAuliffe <mcauliff <@t> umdnj.edu>
Subject: Re: [Histonet] Problems with c-fos immediate early gene
labelling
To: komo <@t> bu.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <406080E6.2080605 <@t> umdnj.edu>
Content-Type: text/plain; format=flowed; charset=ISO-8859-1
Hi Ron:
komo <@t> bu.edu wrote:
>I'm hoping I could get some helpful advice on what might cause my c-fos
>IHC labelling to work well in one lab, but not my own. I am running the
>protocol on rat brains using a hydrogen peroxide/methanol step for
>endogenous peroxidase blocking, a normal goat serum blocking step, a
>c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated
>secondary, a streptavidin horeradish peroxidase, and a nickle enhanced
>DAB reaction.
>
>Unfortunately, although I have all of the exact same antibodies, chemicals,
and concentrations as the lab from which the protocol was developed, the
label is weak and not very specific when I run it at my home lab. In fact, I
even took brain slices from the same rats that I was running at my lab and
ran them at the lab where the protocol was developed, and found that the
c-fos label was beautiful with a low background.
>
When you say you are using the "exact same antibodies, chemicals,
and ..... " do you mean the same bottle of reagent? You are carrying
the bottle or vial of reagent from one lab to another? If not, look at
the batch of antibody, your peroxide, your DAB, etc. If you are storing
antibody in a freezer at home it is being subjected to freeze-thaw
cycles (unless you have a very old freezer that does not defrost itself
on a timer). If your bottle of peroxide is more than a few months old it
may be in the process of turning into water, peroxide does that. I have
had DAB work one day and fail the next.
Since you have found that the brain slices stain well in the "other
lab", the problem must lie with your home lab. Since bleach reacts with
DAB I would avoid using staining dishes treated with bleach.
Geoff
>Does anyone has any suggestions as to why this might be? One person
>at the other lab suggested that possibly the wells that we run the tissue
in
>were not being cleaned properly (we soak them in bleach while the other
>lab uses just pex). Could this have any effect?
>
>Thanks for any advice you can provide!!
>
>Rob Komorowski
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029
mcauliff <@t> umdnj.edu
**********************************************
------------------------------
Message: 17
Date: Tue, 23 Mar 2004 10:46:32 -0500
From: "McAloose, Dee" <dmcaloose <@t> wcs.org>
Subject: [Histonet] FW: Job opening - Wildlife Conservation Society
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6D3 <@t> WOLF.wcs.org>
Content-Type: text/plain; charset="iso-8859-1"
> Hello,
>
> I'd like to announce an immediate opening for a full-time histotechnician
in the pathology department at the Wildlife Conservation Society located at
the Bronx Zoo. We provide the diagnostic pathology services to one of the
largest zoological collections in the country. Applications are being
accepted until May 15, 2004 or until the position is filled. Please contact
Ms. Tawanda Williams (Human Resources; email: twilliams <@t> wcs.org) if you're
interested in this exciting opportunity and in becoming part of our team!
>
> Thanks for listening and we look forward to hearing from you.
>
> D McAloose, VMD, Dipl ACVP
> Head, Department of Pathology
> Wildlife Conservation Society
> Bronx, NY 10464
> (phone) 718-220-7105
> (fax) 718-220-7126
> dmcaloose <@t> wcs.org
>
>
------------------------------
Message: 18
Date: Tue, 23 Mar 2004 10:21:54 -0600
From: "Jan Shivers" <shive003 <@t> umn.edu>
Subject: [Histonet] IHC on blood and cytology smears (ICC)
To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <006301c410f2$f45d1ed0$78065486 <@t> vdl220FAC>
Content-Type: text/plain; charset="iso-8859-1"
Could someone please send me a protocol for performing IHC/ICC on smears (I
normally only do FFPE slides)?
I'd like info mostly on if the slides have to be coated/charged , smears
air-dried or fixed (and in what) for how long, and if you do any
modification of the immuno protocol, such as blocking endogenous peroxidase
later in the procedure (and with what concentration of H2O2), and/or
changing incubation times/dilutions.
I've done smears on very rare occasions, but have had trouble getting the
cells to remain on the uncoated slides that the smears are being made on.
Thank you very much in advance.
Jan Shivers
Univ of Minnesota Vet Diag Lab
shive003 <@t> tc.umn.edu
------------------------------
Message: 19
Date: Tue, 23 Mar 2004 12:02:46 -0500
From: Luis Chiriboga <Luis.Chiriboga <@t> med.nyu.edu>
Subject: [Histonet] NYSHS 2004 Annual Meeting
To: Histonet <histonet <@t> pathology.swmed.edu>
Message-ID: <KFEIIJOCLBABEKFDAEHPGEMHDNAA.Luis.Chiriboga <@t> med.nyu.edu>
Content-Type: text/plain; charset="iso-8859-1"
The 2004 New York State Histotechnology Society annual meeting will be held
at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April
30th through Saturday May 1st. This years meeting theme is "Histology is no
mystery: Let New York State solve your problems". An e-mail mini program is
available upon request by replying to this message. For a program and
registration packet, please contact Judy LaDuc at 518-897-2247 or
jaladuc <@t> capital.net
We hope to see you there.
------------------------------
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End of Histonet Digest, Vol 4, Issue 27
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