[Histonet] PECAM-1 (sc-1506R) staining

[Victor Wong]

Dear all,
 
Thank you for all the professional suggestion of the previous IHC staining and 
finally I got some positive results.
 
Then comes problem on the PECAM-1 IHC.  I am using the Santa Cruz sc-1506R (it 
is a anti-RAT antibody from RABBIT).  I stain decalcified rat spine paraffin 
sections, with following steps:
 
antigen retrieval (~95C MW, DAKO AR solution pH6 for 20 minutes);
DAKO's Dual Enzyme block for 10 minutes;
3% normal goat serum for 20 minutes;
1:50 primary antibody at 4C for overnight;
DAKO's Envision anti-rabbit kit for 30 minutes;
DAKO's DAB solution to develop
 
I used rat liver and kidney as positive controls.  I got background staining on 
hepatocytes but none staining on vessels.  On kidney section, nucleus and 
lining within tubules and basement membrane of glomerulus were also stained but 
not that bad as liver.  I cannot say it was totally non-specific as some 
nucleus were not stained.
 
I think PECAM-1 IHC needs more specific conditons.  Could any histoneters have 
any suggestion?
 
Many thanks in advance.
 
Victor



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Re: [Histonet] ? on Gulf oil spill

[Lesley Weston]

Oil Red O might work, since it stains lipids.

Lesley Weston.


On 2010-07-28, at 7:26 AM, jstaruk wrote:

> Hi all,
> 
> I have a customer who wants to study the amounts of oil accumulation in the
> gills of fish from the Gulf of Mexico.  Does anyone have any suggestions as
> to what techniques might stain crude oil? 
> 
> Thanks
> 
> Jim
> 
> ___
> James E. Staruk HT(ASCP)
> www.masshistology.com
>   www.nehorselabs.com
> 
> 
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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Re: [Histonet] Refrigerate formalin?

[Mark Ray]

It is usually recommended that 10% Formalin Fixative be stored at 
temperatures above about 10C to minimize polymerization of 
Formaldehyde.  You might confirm this with the manufacturer of your 
Formalin Fixative.  The label may also say something about this.



Breeden, Sara wrote:

We're getting ready to move into our new building (YAHOO!) in early
September - finally.  In planning where we will store our cases for the
required 6-week retention time, I have proposed that the tissues (in 10%
NBF) be shelved in the walk-in cooler (4 degrees C).   Is there any
reason tissues-in-formalin could NOT be stored refrigerated for that
length of time?  It is extremely rare that we are required to pull and
recut wet tissue, but that possibility always exists.  Thanks, everyone.

 


Sally Breeden, HT(ASCP)

Veterinary Diagnostic Services

New Mexico Department of Agriculture

700 Camino de Salud NE

Albuquerque, NM  87108

505-841-2576

 


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[Histonet] REFRIG FORMALIN

[Tench, Bill]

You must have a heck of a lot of space in the frig.  I am envious. I
don't think refrigerating your specimens would create any problems.
Everyone i know stores them in some inconvenient corner of a morgue at
room temperature.
 
Bill Tench
Associate Dir. Laboratory Services
Chief, Cytology Services
Palomar Medical Center
555 E. Valley Parkway
Escondido, California  92025
bill.te...@pph.org
Voice: 760- 739-3037
Fax: 760-739-2604
 

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Re: [Histonet] Refrigerate formalin?

[Rene J Buesa]

There is nothing against cooling formalin fixed tissues, but my question is WHY?
I see it as a waste of refrigerated space. 
On the other hand also, IF by any change there is a formalin spill inside the 
walk-in cooler room, that will be a royal mess to decontaminate.
René J.

--- On Wed, 7/28/10, Breeden, Sara  wrote:


From: Breeden, Sara 
Subject: [Histonet] Refrigerate formalin?
To: histonet@lists.utsouthwestern.edu
Date: Wednesday, July 28, 2010, 4:41 PM


We're getting ready to move into our new building (YAHOO!) in early
September - finally.  In planning where we will store our cases for the
required 6-week retention time, I have proposed that the tissues (in 10%
NBF) be shelved in the walk-in cooler (4 degrees C).   Is there any
reason tissues-in-formalin could NOT be stored refrigerated for that
length of time?  It is extremely rare that we are required to pull and
recut wet tissue, but that possibility always exists.  Thanks, everyone.



Sally Breeden, HT(ASCP)

Veterinary Diagnostic Services

New Mexico Department of Agriculture

700 Camino de Salud NE

Albuquerque, NM  87108

505-841-2576



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[Histonet] Refrigerate formalin?

[Breeden, Sara]

We're getting ready to move into our new building (YAHOO!) in early
September - finally.  In planning where we will store our cases for the
required 6-week retention time, I have proposed that the tissues (in 10%
NBF) be shelved in the walk-in cooler (4 degrees C).   Is there any
reason tissues-in-formalin could NOT be stored refrigerated for that
length of time?  It is extremely rare that we are required to pull and
recut wet tissue, but that possibility always exists.  Thanks, everyone.

 

Sally Breeden, HT(ASCP)

Veterinary Diagnostic Services

New Mexico Department of Agriculture

700 Camino de Salud NE

Albuquerque, NM  87108

505-841-2576

 

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[Histonet] dissection aide

[Tench, Bill]

I don't see much advantage of this over plain old Carnoy's fixative,
other than missing the wiff of choroform.  Ethanol should not be a
problem with IHC, but as you said, revalidation is required.  The magic
number of nodes is 12.
 
Bill Tench
Associate Dir. Laboratory Services
Chief, Cytology Services
Palomar Medical Center
555 E. Valley Parkway
Escondido, California  92025
bill.te...@pph.org
Voice: 760- 739-3037
Fax: 760-739-2604
 

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[Histonet] Sakura VIP6 update

[Cynthia Pyse]

Thanks everyone for your responses. Sakura has scheduled an upgrade of my
rotary and gate valve. I hope this is the answer I need.

Cindy

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[Histonet] RE: Histonet Digest, Vol 80, Issue 33

[Joanne Clark]

We use a similar product called Dissect Aid, from Decal Corp.  IHC staining has 
never been an issue; we never run IHC on the lymph nodes from colon cancer 
cases.  We usually do the IHC on the tumor itself which is fixed in formalin.  
This kind of product is very useful in helping to find all those pesky little 
nodes that like to hide in the fat by turning them white.  Cancer protocols 
require that at least 13 nodes be submitted.  As histo supervisor, I also do 
the grossing in my facility, so I understand why your PA is interested in this 
kind of product (it can be difficult to come up with this magic number of 13 
without it).  However, if you do run IHC on the nodes from these cases, you 
would indeed have to revalidate your markers with this as your primary fixative 
or as a post fixative.

Joanne Clark, HT, MLT
Histology Supervisor
Pathology Consultants of New Mexico
Roswell, NM


-

Message: 2
Date: Tue, 27 Jul 2010 15:03:33 -0300
From: "Hayes, Randi (HorizonNB)" 
Subject: [Histonet] Using GEWF solution and IHC staining
To: 
Message-ID: 
Content-Type: text/plain; charset="iso-8859-1"

At a recent conference, our PA learned of using GEWF (glacial acetic acid, 
ethanol, distilled water, 40% formaldehyde) solution as an aid for Lymph Node 
retrieval in Colorectal Cancer resections.  Although a good idea, I'm wondering 
how "safe" it is to use when staining for IHC.  Does anyone have much 
experience with this or know of a study (studies) that have been done to verify 
that the ethanol is not destroying antigen sites?  We're a little 
concerned..


Randi Hayes, MLT
Histology Supervisor / Superviseur d'Histologie
Horizon Health Network / Réseau de santé Horizon
(506) 860-2157
randi.ha...@horizonnb.ca  
www.HorizonNB.ca  





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[Histonet] Gordon and Sweets retic stain

[Johnson, Nacaela]

 
Hello! 

I am having a problem with my tissue washing off of the slides during
the retic stain. The majority of the tissue is falling off during the
Working silver solution step because of the alkalinity of the solution.
The tissue is bone marrow and I use Halt in my water bath. My thoughts
are to add a mild acid to the working solution to bring the pH down, but
I am not sure of what the effects are on the solution. I do not want the
silver to not impregnate. Has anyone had this problem before and found a
solution? 

 
 
Thanks,
 
Nacaela Johnson
Histology Technician
KCCC Pathology
12000 110th St., Ste. 400
Overland Park, KS 66210
Office:  913-234-0576
Fax:  913-433-7639
Email:  nacaela.john...@usoncology.com
 
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[Histonet] Sakura VIP6

[Cynthia Pyse]

Help!

I am having a problem with me VIP6 tissue processor. About every 4 to 6
weeks the processor has a error code that position 2 which is 80% has not
pumped in fully in the allotted time. Sakura answers to this problem is
either I need to use their formalin or the alcohol in the second station is
too high a concentration. I have never had this problem before in any
processor I have used. I still use the Shandon excelsiour tissue processor
and use the same protocol in both machines with no problems in the Shandon.
Sakura thinks it is a build-up of precipitating salts that is causing the
problem but there was no sign of this when the service tech checked the
rotary valve. After every process we not only flush with ETOH and xylene but
also run a warm water flush from the first station which is formalin. I use
a different bottle for the warm water but do use the same valve from the
formalin bottle. When we have completed 7 runs we run the 3 warm water flush
Sakura recommends. Even the service tech remarked that I was doing more than
they recommend.

 I am at a loss as to what it could be. My only conclusion is that maybe the
valve in station 2 intermittently collapse or sticks for some reason. I
wanted to see if anyone either is having the same problem or if someone has
a thought of what the problem could be. I did purchase a refurbished
machine. I would appreciate any thoughts, ideas, or even guesses that anyone
has. 

Thanks in advance for any help.

 

Cindy Pyse, CLT, HT (ASCP)

Laboratory/Histology Supervisor

X-Cell Laboratories

716-250-9235

e-mail cp...@x-celllab.com

 

 



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RE: [Histonet] ? on Gulf oil spill

[Roberta Horner]

I did an Oil Red O on beaver tissues from a place there was a diesel spill and 
it worked out pretty well.
Roberta Horner HT/HTL
Penn State University
Animal Diagnostic Lab

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of jstaruk
Sent: Wednesday, July 28, 2010 10:26 AM
Cc: histonet@lists.utsouthwestern.edu
Subject: [Histonet] ? on Gulf oil spill

Hi all,

I have a customer who wants to study the amounts of oil accumulation in the
gills of fish from the Gulf of Mexico.  Does anyone have any suggestions as
to what techniques might stain crude oil? 

Thanks

Jim

___
James E. Staruk HT(ASCP)
 www.masshistology.com
   www.nehorselabs.com


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[Histonet] ? on Gulf oil spill

[jstaruk]

Hi all,

I have a customer who wants to study the amounts of oil accumulation in the
gills of fish from the Gulf of Mexico.  Does anyone have any suggestions as
to what techniques might stain crude oil? 

Thanks

Jim

___
James E. Staruk HT(ASCP)
 www.masshistology.com
   www.nehorselabs.com


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[Histonet] Histology Tech Position In Irvine CA.

[Masterson_John]

Allergan Inc. in Irvine California is looking to fill a position in our DSE 
Pathology Dept. If interested please see details on the Allergan web site.  

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[Histonet] RES: Histonet Digest, Vol 80, Issue 32

[Thalita]

Vanusa,

Não tenho permissão para mexer no Igecor e não entendi o que deveria ser
arrumado.

Att,


Thalita Ferraz
Produção
+55 12 3203-0612 (DIRETO)
+55 12 3203-0633 (SAC)
www.cipax.com.br
thal...@cipax.com.br


-Mensagem original-
De: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Em nome de
histonet-requ...@lists.utsouthwestern.edu
Enviada em: terça-feira, 27 de julho de 2010 14:07
Para: histonet@lists.utsouthwestern.edu
Assunto: Histonet Digest, Vol 80, Issue 32

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Today's Topics:

   1. Re:  [Sectioning]  Brain Tissue (Nancy Herman)
   2. Re: Powdered reagent expiration dates (Pat Laurie)
   3. Tissue processing protocol for Hamster tissues (abijag )


--

Message: 1
Date: Tue, 27 Jul 2010 11:25:54 -0400
From: "Nancy Herman" 
Subject: [Histonet] Re:  [Sectioning]  Brain Tissue
To: ,"Margaret Sherwood"

Message-ID: <4c4ea622.6deb.00d...@inspection.gc.ca>
Content-Type: text/plain; charset=US-ASCII

We keep our water bath at 40-41 C.  Sometimes we add some 70% ethanol to the
water bath and this helps to get the wrinkles out (especially for cerebellum
and mice brain).  Most of our tissue is bovine brain but we deal with
multiple other species including mice.  We use McCormick Paraplast Plus
embedding Medium from Fisher.

Nancy Herman
Canadian Food Inspection Agency - Lethbridge Laboratory

>>> "Sherwood, Margaret "  2010/07/27 9:14 am >>>
To All:
 
We have been experiencing problems sectioning mouse brain.  The sectioning
is
fine, but we have problems in the water bath.  At 45 degrees C (peel-a-way
paraffin-Polysciences), the sections break apart but we don't get wrinkles.
With paraplast extra, we use 36 degrees C, but the sections are wrinkled and
disintegrate.   
 
I would appreciate hearing from listers as to what you do:  what temperature
is
your water bath, what embedding media do you use, etc?  Any help would be
appreciated!
 
Thanks!
Peggy
 
Peggy Sherwood
Lab Associate, Photopathology 
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherw...@partners.org 
 


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Message: 2
Date: Tue, 27 Jul 2010 09:00:20 -0700
From: Pat Laurie 
Subject: Re: [Histonet] Powdered reagent expiration dates
To: lpw...@sbcglobal.net
Cc: Histonet@lists.utsouthwestern.edu
Message-ID:

Content-Type: text/plain; charset=ISO-8859-1

Everyone,

Thanks for your help, based off your explanations, statements and citations,
we will attempt to protest this and have it expunged.



On Mon, Jul 26, 2010 at 4:41 PM, Lee & Peggy Wenk
wrote:

> See if you can get the following article. Biotech Histochem is published
by
> the Biological Stain Commission. http://www.biologicalstaincommission.org/
>
> Biotech Histochem. 2009 Feb;84(1):11-5.
>
> Stain and dye stability over a 30-year period: a comparison of certified
> dye
> powders by the Biological Stain Commission.
> Penney DP, Frank M, Fagan C, Willis C.
>
> Biological Stain Commission, Department of Pathology & Laboratory
Medicine,
> University of Rochester Medical Center, Rochester, New York 14642-0001,
> USA.
> david_pen...@urmc.rochester.edu
>
> Abstract
> The Biological Stain Commission (BSC) Assay Laboratory has received
> numerous
> inquiries during the past several years regarding the long-term stability
> of
> stain and dye powders, particularly since packaging requirements call for
> expiration dates on reagents. We have conducted a study to examine the
> long-term stability of selected dye powders. We used the standard
> procedures
> of the BSC for testing biological stains for certification to give an
> indication of the long-term chemical stability as well as staining
> p