[Histonet] smears for IHC

Wed Mar 24 16:22:22 CST 2004

A point worth mentioning here is that fixation of smears can be
accomplished using this vapor method with a variety of fixatives,
formaldehyde, glutaraldehyde, acetone, ethanol, osmium tetroxide etc.
The major advantage of  vapor fixation is that the tissue is not exposed
to the vehicle containing the fixative. Because of this thin layers such
as smears tend to stay on the slides better than if placing these in a
solution of the fixative. It also eliminates the need to worry about
osmolarity, concentration and pH. It is however, only useful for thin
layers, cryostat sections etc.
Barry

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Dorothy.L.Webb <@t> HealthPartners.Com
Sent: Wednesday, March 24, 2004 2:37 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] smears for IHC

Whenever we do IHC on smears, we fix the smears in equal parts of
hydrogen peroxide and methyl alcohol for 15 minutes.  Another method
that can be used is to place smears in a coplin jar and place
37%formaldehyde soaked gauze in the lid.  Cover the smears for 20
minutes.  Hope this helps!  Dorothy Webb at Regions Hospital, St. Paul,
Minn,

-----Original Message-----
From: histonet-request <@t> lists.utsouthwestern.edu
[mailto:histonet-request <@t> lists.utsouthwestern.edu]
Sent: Tuesday, March 23, 2004 12:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 4, Issue 27

Send Histonet mailing list submissions to
	histonet <@t> lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
	histonet-request <@t> lists.utsouthwestern.edu

You can reach the person managing the list at
	histonet-owner <@t> lists.utsouthwestern.edu

When replying, please edit your Subject line so it is more specific than
"Re: Contents of Histonet digest..."

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

CONFIDENTIALITY NOTICE: 
This email transmission and any documents, files, 
or previous e-mail messages attached to it are 
confidential and intended solely for the use of the 
individual or entity to whom they are addressed. 
If you are not the intended recipient, or a person 
responsible for delivering it to the intended recipient, 
you are hereby notified that any further review, 
disclosure, copying, dissemination, distribution, or 
use of any of the information contained in or attached 
to this e-mail transmission is strictly prohibited. 
If you have received this message in error, please 
notify the sender immediately by e-mail, discard 
any paper copies, and delete all electronic files 
of the message. If you are unable to contact the 
sender or you are not sure as to whether you 
are the intended recipient, please e-mail 
ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777. 

------------------------------

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

CONFIDENTIALITY NOTICE: 
This email transmission and any documents, files, 
or previous e-mail messages attached to it are 
confidential and intended solely for the use of the 
individual or entity to whom they are addressed. 
If you are not the intended recipient, or a person 
responsible for delivering it to the intended recipient, 
you are hereby notified that any further review, 
disclosure, copying, dissemination, distribution, or 
use of any of the information contained in or attached 
to this e-mail transmission is strictly prohibited. 
If you have received this message in error, please 
notify the sender immediately by e-mail, discard 
any paper copies, and delete all electronic files 
of the message. If you are unable to contact the 
sender or you are not sure as to whether you 
are the intended recipient, please e-mail 
ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777. 

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

------------------------------

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
	confidential and intended solely for the use of the
	individual or entity to whom they are addressed.
	If you are not the intended recipient, or a person
	responsible for delivering it to the intended recipient,
	you are hereby notified that any further review,
	disclosure, copying, dissemination, distribution, or
	use of any of the information contained in or attached
	to this e-mail transmission is strictly prohibited.
	If you have received this message in error, please
	notify the sender immediately by e-mail, discard
	any paper copies, and delete all electronic files
	of the message. If you are unable to contact the
	sender or you are not sure as to whether you
	are the intended recipient, please e-mail
	ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777.

	_______________________________________________
	Histonet mailing list
	Histonet <@t> lists.utsouthwestern.edu
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet

------------------------------

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.co
m>

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. _______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

**********************************************************************
This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient, please 
delete it and notify the sender.

Views expressed in this message and any attachments are those 
of the individual sender, and are not necessarily the views of The
Children's Hospital at Westmead

This footnote also confirms that this email message has been 
virus scanned and although no computer viruses were detected, the
Childrens Hospital at Westmead accepts no liability for any 
consequential damage resulting from email containing computer 
viruses.
**********************************************************************

------------------------------

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 
Disclaimer: The information contained in this message and/or any
attachments(s) may be of a private and confidential nature, and is
intended solely for the attention of the addressee. If you have received
this message in error or feel you should not have been the intended
recipient, please return it and any attachments to the sender
immediately. All messages relating to this communication should then be
deleted from your system. Unauthorised usage, copying, disclosure or
alteration of this message and/or attachment(s) is strictly prohibited.
Barking, Havering and Redbridge Hospitals NHS Trust will not be held
responsible for any direct or indirect damages which may arise from
alteration of this message or any attachment(s), by a third party or
resulting from the transmission of a virus.

-----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
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

CONFIDENTIALITY NOTICE: 
This email transmission and any documents, files, 
or previous e-mail messages attached to it are 
confidential and intended solely for the use of the 
individual or entity to whom they are addressed. 
If you are not the intended recipient, or a person 
responsible for delivering it to the intended recipient, 
you are hereby notified that any further review, 
disclosure, copying, dissemination, distribution, or 
use of any of the information contained in or attached 
to this e-mail transmission is strictly prohibited. 
If you have received this message in error, please 
notify the sender immediately by e-mail, discard 
any paper copies, and delete all electronic files 
of the message. If you are unable to contact the 
sender or you are not sure as to whether you 
are the intended recipient, please e-mail 
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

_______________________________________________
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

CONFIDENTIALITY NOTICE: 
This email transmission and any documents, files, 
or previous e-mail messages attached to it are 
confidential and intended solely for the use of the 
individual or entity to whom they are addressed. 
If you are not the intended recipient, or a person 
responsible for delivering it to the intended recipient, 
you are hereby notified that any further review, 
disclosure, copying, dissemination, distribution, or 
use of any of the information contained in or attached 
to this e-mail transmission is strictly prohibited. 
If you have received this message in error, please 
notify the sender immediately by e-mail, discard 
any paper copies, and delete all electronic files 
of the message. If you are unable to contact the 
sender or you are not sure as to whether you 
are the intended recipient, please e-mail 
ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777. 

------------------------------

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.

------------------------------

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

End of Histonet Digest, Vol 4, Issue 27
***************************************

________________________________________

This e-mail and any files transmitted with it are confidential and are 
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please
be advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is
strictly prohibited.

If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will
be reimbursed for reasonable costs incurred in notifying us.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet