[Histonet] Formaline free fixative

Thu Nov 8 16:16:08 CST 2007

Dear Bob,
I know indeed quite a number of glyoxal based fixatives. Many labs have
tested them over here. Products were FineFix, GlyoFix, amongst others.
However, most of time morphology results were OK but IHC was bad. It looks
like glyoxal tends to fix too much. 
As for MSDS, yes, they are required as well in France and I have noticed
that allowed ppm in lab air is even less that with formalin !
Do you have yourself any experience with glyoxal based fixative ?
Best regards
Pierre

-----Message d'origine-----
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Envoyé : jeudi 8 novembre 2007 19:07
À : histonet <@t> lists.utsouthwestern.edu
Objet : Histonet Digest, Vol 48, Issue 11

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Today's Topics:

   1. mouse histology atlas (Helen E Johnson)
   2. Re: Away from Office (arnie.jimenez <@t> vel-lab.com)
   3. Short processing schedule-suggestions (Greg Dobbin)
   4. Re: Formaline-free fixative (Robert Richmond)
   5. B5 substitute (Joanne Mauger)
   6. RE: Artefacts (Rene J Buesa)
   7. Re: Short processing schedule-suggestions (Rene J Buesa)
   8. Clone  for Prog. Receptor (Drew Sally A.)
   9. Re: Artefacts (John Kiernan)
  10. RE: Artefacts (Marshall Terry Dr,	Consultant Histopathologist)
  11. Re: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO)
  12. RE: Artefacts (Weems, Joyce)
  13. RE: B5 substitute (Weems, Joyce)
  14. Re: B5 substitute (Matt Bancroft)
  15. RE: Artefacts (Marshall Terry Dr,	Consultant Histopathologist)
  16. Re: B5 substitute (Richard Cartun)
  17. Re: Clone  for Prog. Receptor (Richard Cartun)
  18. RE: problem in dual staining of CD4 and FOXP3 (Tarango, Mark)
  19. RE: B5 substitute (Tarango, Mark)
  20. HT position wanted (Steven Coakley)
  21. RE: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO)

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

Message: 1
Date: Thu, 8 Nov 2007 09:22:44 -0500
From: Helen E Johnson <hej01 <@t> health.state.ny.us>
Subject: [Histonet] mouse histology atlas
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:

<OF658DCF06.47B7C4B1-ON8525738D.004ED347-8525738D.004EFE75 <@t> notes.health.stat
e.ny.us>

Content-Type: text/plain; charset=US-ASCII

Hi Histonetters,
     Does anyone know of a good mouse histology atlas?
                              Helen Johnson  (hej01 <@t> health.state.ny.us)

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

Message: 2
Date: 8 Nov 2007 08:23:20 -0600
From: arnie.jimenez <@t> vel-lab.com
Subject: [Histonet] Re: Away from Office
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20071108142320.15276.qmail <@t> plesk5.hostgator.com>
Content-Type: text/plain; charset="UTF-8"

Thank you for contacting Vel-Lab Research. We will be out of the lab
starting Nov. 7th and will return on Nov. 12th. We will check e-mail
regularly but will be unable to respond. Please do not mail any packages to
our lab until Nov. 12th.

Regards,

Arnie Jimenez
Vel-Lab Research

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

Message: 3
Date: Thu, 08 Nov 2007 10:59:21 -0400
From: "Greg Dobbin" <gvdobbin <@t> ihis.org>
Subject: [Histonet] Short processing schedule-suggestions
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s732ec1e.013 <@t> ihis.org>
Content-Type: text/plain; charset=US-ASCII

Hello All,
I would like to get your suggestions for a processing schedule for small
biopsies (cores, GI's, other endoscopics). I want to process these little
guys separate from everything else to hopefully improve section quality and
perhaps staining quality. 

I don't want the absolute shortest possible schedule. We have a newer
VIP5 for our main processor (doing everything on one schedule currently) and
a much older VIP sitting idle as a backup that I plan to start using for the
biopsies. We  process overnight so I have more than enough time.
I will just delay the biopsy processing so that both processors are finished
at essentially the same time. 

What I need is a shorter schedule for the biopsies so that the detrimental
affects of xylene and alcohol are minimized (and yes, we still use xylene!
And in case it matters schedule-wise, we use 10% NBFS as well).

Thanks in advance.
Greg 

Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE    C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385

There is some merit in doing the right thing rather badly, but absolutely
none in doing the wrong thing excellently!

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

Message: 4
Date: Thu, 8 Nov 2007 10:08:27 -0600
From: "Robert Richmond" <RSRICHMOND <@t> aol.com>
Subject: [Histonet] Re: Formaline-free fixative
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<abea52a60711080808n2186fd70ncda003c722b16ab2 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Pierre Chaumat (where are you, Pierre?) asks:

>>Has anyone tested alternative solution to formaline fixation ? Has 
>>anyone
switched to a new tissue fixative ? - I would like to share some
experience.<<

About the only alternative "fixative" that actually is a fixative is
glyoxal, available under numerous trade names. Its fixative properties are
somewhat different from those of formaldehyde. In particular, immunostains
may require adjustment, and comparison with formaldehyde fixed tissue from
the same case. In the USA, the breast cancer immunostains are required by
the federal govenrment to be done on formaldehyde fixed tissue.

Do not use any trade-named fixative whose composition is secret.
Sometimes fairly adequate information can be obtained from the Materials
Safety Data Sheet (MSDS), in the USA. Does France require similar
documentation of hazardous materials? If so, what is this document called,
and what is a good Web site to look some of them up?
(I can read French.)

Bob Richmond
Samurai Pathologist
Knoxville, Tennessee

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

Message: 5
Date: Thu, 08 Nov 2007 11:50:43 -0500
From: "Joanne Mauger" <mauger <@t> email.chop.edu>
Subject: [Histonet] B5 substitute
To: <gvdobbin <@t> ihis.org>,<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s732f82a.014 <@t> email.chop.edu>
Content-Type: text/plain; charset=US-ASCII

Hi ,

What are you using to replace B5 fixative with mercury? I want the best for
hematopoetic tissues, and for immunostains.

Thanks,

Jo Mauger

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

Message: 6
Date: Thu, 8 Nov 2007 08:59:40 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: RE: [Histonet] Artefacts
To: Kemlo Rogerson <Kemlo.Rogerson <@t> waht.swest.nhs.uk>,	Toxicology
	<dis.tox <@t> suven.com>, histonet <@t> lists.utsouthwestern.edu
Message-ID: <222219.92551.qm <@t> web61219.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

and most likely the first!
Reni J.

Kemlo Rogerson <Kemlo.Rogerson <@t> waht.swest.nhs.uk> wrote:  Ah the spotty H&E
problem again.

1) Inadequate dewaxing.
2) Inadequate fixation.
3) Inadequate processing.

Not necessarily in that order.

Kemlo Rogerson
Pathology Manager
DD 01934 647057 or extension 3311
Mob 07749 754194; Pager 07659 597107;
No snowflake in an avalanche ever feels responsible. --George Burns 

This e-mail is confidential and privileged. If you are not the intended
recipient please accept my apologies; please do not disclose, copy or
distribute information in this e-mail or take any action in reliance on its
contents: to do so is strictly prohibited and may be unlawful.
Please inform me that this message has gone astray before deleting it.
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------------------------------

Message: 7
Date: Thu, 8 Nov 2007 09:03:38 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Short processing schedule-suggestions
To: Greg Dobbin <gvdobbin <@t> ihis.org>, Histonet <@t> lists.utsouthwestern.edu
Message-ID: <462982.42437.qm <@t> web61214.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

>From my experience in processing, the most important thing is to maintain
dehydration time to clearing time to infiltration time in constant
proportions once you find a good protocol.
  If you "long" protocol works well for you, reduce the time in each step in
a way that at the end will be completed in the time you need, BUT preserving
the time proportions you have now.
  Reni J.

Greg Dobbin <gvdobbin <@t> ihis.org> wrote:
  Hello All,
I would like to get your suggestions for a processing schedule for small
biopsies (cores, GI's, other endoscopics). I want to process these little
guys separate from everything else to hopefully improve section quality and
perhaps staining quality. 

I don't want the absolute shortest possible schedule. We have a newer
VIP5 for our main processor (doing everything on one schedule currently) and
a much older VIP sitting idle as a backup that I plan to start using for the
biopsies. We process overnight so I have more than enough time.
I will just delay the biopsy processing so that both processors are finished
at essentially the same time. 

What I need is a shorter schedule for the biopsies so that the detrimental
affects of xylene and alcohol are minimized (and yes, we still use xylene!
And in case it matters schedule-wise, we use 10% NBFS as well).

Thanks in advance.
Greg 

Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385

There is some merit in doing the right thing rather badly, but absolutely
none in doing the wrong thing excellently!

Statement of Confidentiality
This message (including attachments) may contain confidential or privileged
information intended for a specific individual or organization. If you have
received this communication in error, please notify the sender immediately.
If you are not the intended recipient, you are not authorized to use,
disclose, distribute, copy, print or rely on this email, and should promptly
delete this email from your entire computer system. 

D?claration de confidentialit?
Le pr?sent message (y compris les annexes) peut contenir des renseignements
confidentiels ayant pour objet une personne ou un organisme particulier. Si
vous avez re?u la pr?sente communication par erreur, veuillez en informer
l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous
n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce
courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement
de votre syst?me informatique.

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

 __________________________________________________
Do You Yahoo!?
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------------------------------

Message: 8
Date: Thu, 8 Nov 2007 11:07:25 -0600
From: "Drew Sally A." <SDrew <@t> uwhealth.org>
Subject: [Histonet] Clone  for Prog. Receptor
To: "Histonet" <histonet <@t> pathology.swmed.edu>
Message-ID:

<AF782AEBFF55684781F61CEFB8BFC7310AD57F <@t> uwhis-xchng3.uwhis.hosp.wisc.edu>

Content-Type: text/plain;	charset="us-ascii"

We currently are using the 1A6 clone for our progesterone receptor(PR) IHC.
We were wondering how other people felt about rabbit monoclonal PR
antibodies.
Does anyone have strong preferences or experiences with other PR
clones-mouse or rabbit-that they'd like to share? 
   Thank you...!

Sally Ann Drew, MT(ASCP)
IHC/ISH Laboratory
University of Wisconsin Hosp. & Clinics
Madison, WI 53792
(608)265-6596

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

Message: 9
Date: Thu, 08 Nov 2007 12:10:47 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] Artefacts
To: Toxicology <dis.tox <@t> suven.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <de2ea3e92e498.4732fcc7 <@t> uwo.ca>
Content-Type: text/plain; charset=us-ascii

Dear Toxicology <dis.tox <@t> suven.com>

You need to explain your problem more clearly. Please include
histonet <@t> lists.utsouthwestern.edu in your replies. We all want to share our
knowledge.

What tissue are you staining? How was it fixed?  Are you staining dewaxed
and hydrated paraffin sections?  Cryostat sections?  

Which haemalum & eosin method did you use?  Does your lab have a book with
instructions for H&E staining?  If not, why not?

Are the "faint stained spots on sections" blue, purple, pink or red? Where
are these spots?  

John Kiernan
Anatomy, UWO
London, Canada.
===
----- Original Message -----
From: Toxicology <dis.tox <@t> suven.com>
Date: Thursday, November 8, 2007 1:36
Subject: [Histonet] Artefacts
To: histonet <@t> lists.utsouthwestern.edu

> Dear all,
> I am facing a problem in Hematoxylin and Eosin staining. There are 
> several faint stained spots on sections and because of that its 
> difficuly to interpret the lesions.  I am unable to sort out the 
> problem. Can any one help me?
>  
> 
> 
> 
> -------------------------Email Disclaimer------------------------
> ---------
> 

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

Message: 10
Date: Thu, 8 Nov 2007 17:13:21 -0000
From: "Marshall Terry Dr,	Consultant Histopathologist"
	<Terry.Marshall <@t> rothgen.nhs.uk>
Subject: RE: [Histonet] Artefacts
To: "John Kiernan" <jkiernan <@t> uwo.ca>, "Toxicology" <dis.tox <@t> suven.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<5C0BED61F529364E86309CADEA63FEF2F3AE4F <@t> TRFT-EX01.xRothGen.nhs.uk>
Content-Type: text/plain;	charset="us-ascii"

Some of these questions should be answered by the photos, but I cannot find
them.

Terry 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John Kiernan
Sent: 08 November 2007 17:11
To: Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Artefacts

Dear Toxicology <dis.tox <@t> suven.com>

You need to explain your problem more clearly. Please include
histonet <@t> lists.utsouthwestern.edu in your replies. We all want to share our
knowledge.

What tissue are you staining? How was it fixed?  Are you staining dewaxed
and hydrated paraffin sections?  Cryostat sections?  

Which haemalum & eosin method did you use?  Does your lab have a book with
instructions for H&E staining?  If not, why not?

Are the "faint stained spots on sections" blue, purple, pink or red?
Where are these spots?  

John Kiernan
Anatomy, UWO
London, Canada.
===
----- Original Message -----
From: Toxicology <dis.tox <@t> suven.com>
Date: Thursday, November 8, 2007 1:36
Subject: [Histonet] Artefacts
To: histonet <@t> lists.utsouthwestern.edu

> Dear all,
> I am facing a problem in Hematoxylin and Eosin staining. There are 
> several faint stained spots on sections and because of that its 
> difficuly to interpret the lesions.  I am unable to sort out the 
> problem. Can any one help me?
>  
> 
> 
> 
> -------------------------Email Disclaimer------------------------
> ---------
> 
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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

Message: 11
Date: Thu, 8 Nov 2007 12:13:22 -0500 (EST)
From: "FU,DONGTAO" <fudo <@t> ufl.edu>
Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<813060655.505211194542002919.JavaMail.osg <@t> osgjas04.cns.ufl.edu>
Content-Type: text/plain; format=flowed; charset=us-ascii

Hello,

  Can anyone give me some suggestions on my case? I know a lot of people
here have a lot of dual staining experiences. I did dual staining on 2
antibodies from different species in the past. They worked well. But using 2
antibodies from same species is my first time try and I met a big problem.
Any suggestions I will be very appreciate.

Ann

On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" <fudo <@t> ufl.edu>
wrote:

> Hi, all
> 
>   Thank you first for giving me some good suggestions on thick-section 
> question I posted last time. Now I met another problem when I did CD4 
> and FOXP3 dual staining using murine fresh frozen spleen. If I did 
> single staining, both antibodies worked very well. However if I did 
> dual staining, I could only get good result from the first antibody. 
> The second one did not work at all(I mean no specific staining). I 
> used CD4 from BD(rat anti-mouse). FOXP3 I used from ebioscience, also 
> rat anti-mouse.
> I think there might be something wrong with my serum block(I used 
> normal rat serum block) before I added secondary antibody. Or any 
> other serum block I need to add to decrease the non-specific binding 
> which I have not done yet. Does anyone can give me some suggestions 
> according to my protocol below? How can I get specific staining of the 
> secondary (primary)antibody?  Thank you,
> 
>   Below is the simple protocol I used for dual staining:
>   1. 2% normal goat serum block 20 min
>   2. 1* antibody CD4 1:750 in diluent O/N 4C
>   3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT
>   4. Serum block:  5% normal rat serum 30 min
>   5. 1* antibody FOXP3 1:100 in diluent 1h RT
>   6. Secondary AF488 Donkey anti-rat in TBS 1h RT
> 
>   Use 1xTBS as wash buffer. Before staining, fix tissue in -20C Aceton 
> for 5 min, then airdry.
> 
> 
> Ann Dongtao Fu MD Ph.D
> Lab Manager
> Molecular Pathology core
> Dept. of Pathology
> University of Florida
> 1600 SW Archer Road
> Gainesville, FL 32610
> Lab Phone: 352-273-7752
> FAX: 352-273-7753
> Rm: D11-50
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 

Ann Dongtao Fu MD, Ph.D
Lab Manager
Dept. of Pathology
Lab phone: 352-273-7752
Lab FAX: 352-273-7755
Lab address: D11-50
PO Box: 100275
1600 SW Archer Road
University of Flodrida
Gainesville, FL 32610

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

Message: 12
Date: Thu, 8 Nov 2007 12:16:04 -0500
From: "Weems, Joyce" <JWEEMS <@t> sjha.org>
Subject: RE: [Histonet] Artefacts
To: "Marshall Terry Dr,	Consultant Histopathologist"
	<Terry.Marshall <@t> rothgen.nhs.uk>, 	"John Kiernan"
<jkiernan <@t> uwo.ca>,
	"Toxicology" <dis.tox <@t> suven.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1CD6831EB9B26D45B0A3EAA79F7EBD32048F41AF <@t> sjhaexc02.sjha.org>
Content-Type: text/plain;  charset="utf-8"

The web site says give it 24 hrs for the photo to post...

Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
404-851-7376 - Phone
404-851-7831 - Fax

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Marshall
Terry Dr,Consultant Histopathologist
Sent: Thursday, November 08, 2007 12:13 PM
To: John Kiernan; Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Artefacts

Some of these questions should be answered by the photos, but I cannot
find them.

Terry 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John
Kiernan
Sent: 08 November 2007 17:11
To: Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Artefacts

Dear Toxicology <dis.tox <@t> suven.com>

You need to explain your problem more clearly. Please include
histonet <@t> lists.utsouthwestern.edu in your replies. We all want to share
our knowledge.

What tissue are you staining? How was it fixed?  Are you staining
dewaxed and hydrated paraffin sections?  Cryostat sections?  

Which haemalum & eosin method did you use?  Does your lab have a book
with instructions for H&E staining?  If not, why not?

Are the "faint stained spots on sections" blue, purple, pink or red?
Where are these spots?  

John Kiernan
Anatomy, UWO
London, Canada.
===
----- Original Message -----
From: Toxicology <dis.tox <@t> suven.com>
Date: Thursday, November 8, 2007 1:36
Subject: [Histonet] Artefacts
To: histonet <@t> lists.utsouthwestern.edu

> Dear all,
> I am facing a problem in Hematoxylin and Eosin staining. There are 
> several faint stained spots on sections and because of that its 
> difficuly to interpret the lesions.  I am unable to sort out the 
> problem. Can any one help me?
>  
> 
> 
> 
> -------------------------Email Disclaimer------------------------
> ---------
> 
_______________________________________________
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 ** The information contained in this message may be
privileged and is confidential information intended for the use of the
addressee listed above. If you are neither the intended recipient nor the
employee or agent responsible for delivering this message to the intended
recipient, you are hereby notified that any disclosure, copying,
distribution or the taking of any action in reliance on the contents of this
information is strictly prohibited. If you have received this communication
in error, please notify us immediately by replying to the message and
deleting it from your computer. Thank you. Saint Joseph's Health System,
Inc.

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

Message: 13
Date: Thu, 8 Nov 2007 12:20:12 -0500
From: "Weems, Joyce" <JWEEMS <@t> sjha.org>
Subject: RE: [Histonet] B5 substitute
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<1CD6831EB9B26D45B0A3EAA79F7EBD32048F41B0 <@t> sjhaexc02.sjha.org>
Content-Type: text/plain;  charset="utf-8"

We use B-Plus from BBC. http://www.bbcus.com/website_search.html

It takes a long time to get used to, especially when they have to compare to
previous mercury fixed cases, but it can be done! Good luck, j

Joyce Weems
Pathology Manager
Saint Joseph's Hospital 
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
404-851-7376 - Phone
404-851-7831 - Fax

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Joanne
Mauger
Sent: Thursday, November 08, 2007 11:51 AM
To: gvdobbin <@t> ihis.org; Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] B5 substitute

Hi ,

What are you using to replace B5 fixative with mercury? I want the best
for hematopoetic tissues, and for immunostains.

Thanks,

Jo Mauger

_______________________________________________
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------------------------------

Message: 14
Date: Thu, 8 Nov 2007 09:24:17 -0800 (PST)
From: Matt Bancroft <mohs76009 <@t> yahoo.com>
Subject: Re: [Histonet] B5 substitute
To: Joanne Mauger <mauger <@t> email.chop.edu>, gvdobbin <@t> ihis.org,
	Histonet <@t> lists.utsouthwestern.edu
Message-ID: <704342.79971.qm <@t> web63407.mail.re1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

B-Plus

Joanne Mauger <mauger <@t> email.chop.edu> wrote:
  Hi ,

What are you using to replace B5 fixative with mercury? I want the best
for hematopoetic tissues, and for immunostains.

Thanks,

Jo Mauger

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Message: 15
Date: Thu, 8 Nov 2007 17:27:01 -0000
From: "Marshall Terry Dr,	Consultant Histopathologist"
	<Terry.Marshall <@t> rothgen.nhs.uk>
Subject: RE: [Histonet] Artefacts
To: "Weems, Joyce" <JWEEMS <@t> sjha.org>, "John Kiernan"
	<jkiernan <@t> uwo.ca>,	"Toxicology" <dis.tox <@t> suven.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<5C0BED61F529364E86309CADEA63FEF2F3AE50 <@t> TRFT-EX01.xRothGen.nhs.uk>
Content-Type: text/plain;	charset="us-ascii"

Yes, I know.
Posters can read that too and wait until the photos appear before they
post the question.

Terry 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Weems,
Joyce
Sent: 08 November 2007 17:16
To: Marshall Terry Dr, Consultant Histopathologist; John Kiernan;
Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Artefacts

The web site says give it 24 hrs for the photo to post...

Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
404-851-7376 - Phone
404-851-7831 - Fax

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Marshall
Terry Dr,Consultant Histopathologist
Sent: Thursday, November 08, 2007 12:13 PM
To: John Kiernan; Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Artefacts

Some of these questions should be answered by the photos, but I cannot
find them.

Terry 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John
Kiernan
Sent: 08 November 2007 17:11
To: Toxicology
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Artefacts

Dear Toxicology <dis.tox <@t> suven.com>

You need to explain your problem more clearly. Please include
histonet <@t> lists.utsouthwestern.edu in your replies. We all want to share
our knowledge.

What tissue are you staining? How was it fixed?  Are you staining
dewaxed and hydrated paraffin sections?  Cryostat sections?  

Which haemalum & eosin method did you use?  Does your lab have a book
with instructions for H&E staining?  If not, why not?

Are the "faint stained spots on sections" blue, purple, pink or red?
Where are these spots?  

John Kiernan
Anatomy, UWO
London, Canada.
===
----- Original Message -----
From: Toxicology <dis.tox <@t> suven.com>
Date: Thursday, November 8, 2007 1:36
Subject: [Histonet] Artefacts
To: histonet <@t> lists.utsouthwestern.edu

> Dear all,
> I am facing a problem in Hematoxylin and Eosin staining. There are 
> several faint stained spots on sections and because of that its 
> difficuly to interpret the lesions.  I am unable to sort out the 
> problem. Can any one help me?
>  
> 
> 
> 
> -------------------------Email Disclaimer------------------------
> ---------
> 
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nor the employee or agent responsible for delivering this message to the
intended recipient, you are hereby notified that any disclosure,
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contents of this information is strictly prohibited. If you have
received this communication in error, please notify us immediately by
replying to the message and deleting it from your computer. Thank you.
Saint Joseph's Health System, Inc.

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

Message: 16
Date: Thu, 08 Nov 2007 12:36:02 -0500
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: Re: [Histonet] B5 substitute
To: "Joanne Mauger" <mauger <@t> email.chop.edu>,<gvdobbin <@t> ihis.org>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <473302B20200007700009021 <@t> gwmail4.harthosp.org>
Content-Type: text/plain; charset=US-ASCII

Formalin (fix well and cut it thin).

Richard

Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> "Joanne Mauger" <mauger <@t> email.chop.edu> 11/08/07 11:50 AM >>>
Hi ,

What are you using to replace B5 fixative with mercury? I want the best
for hematopoetic tissues, and for immunostains.

Thanks,

Jo Mauger

_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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This e-mail message, including any attachments, is for the sole use of the
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all copies of the original message.

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

Message: 17
Date: Thu, 08 Nov 2007 12:40:02 -0500
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: Re: [Histonet] Clone  for Prog. Receptor
To: "Histonet" <histonet <@t> pathology.swmed.edu>,	"Drew Sally A."
	<SDrew <@t> uwhealth.org>
Message-ID: <473303A20200007700009027 <@t> gwmail4.harthosp.org>
Content-Type: text/plain; charset=US-ASCII

We have used clone PgR636 (from Dako) for several years now with excellent
results.

Richard

Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> "Drew Sally A." <SDrew <@t> uwhealth.org> 11/08/07 12:07 PM >>>
We currently are using the 1A6 clone for our progesterone receptor(PR)
IHC.
We were wondering how other people felt about rabbit monoclonal PR
antibodies.
Does anyone have strong preferences or experiences with other PR
clones-mouse or rabbit-that 
they'd like to share? 
   Thank you...!

Sally Ann Drew, MT(ASCP)
IHC/ISH Laboratory
University of Wisconsin Hosp. & Clinics
Madison, WI 53792
(608)265-6596

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all copies of the original message.

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

Message: 18
Date: Thu, 8 Nov 2007 09:45:24 -0800
From: "Tarango, Mark" <mtarango <@t> nvcancer.org>
Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3
To: "FU,DONGTAO" <fudo <@t> ufl.edu>,	Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<5AEC610C1CE02945BD63A395BA763EDE011B6F71 <@t> NVCIEXCH02.NVCI.org>
Content-Type: text/plain; charset=us-ascii

Looking at T-regulatory cells, huh?  Are you trying to stain both in the
same color?  I know Foxp3 is nuclear and CD4 is on the cell membrane,
but two colors might be better.  Your protocol looks incomplete.  

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
FU,DONGTAO
Sent: Thursday, November 08, 2007 9:13 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3

Hello,

  Can anyone give me some suggestions on my case? I know a lot of 
people here have a lot of dual staining experiences. I did dual 
staining on 2 antibodies from different species in the past. They 
worked well. But using 2 antibodies from same species is my first 
time try and I met a big problem. Any suggestions I will be very 
appreciate.

Ann

On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" <fudo <@t> ufl.edu> 
wrote:

> Hi, all
> 
>   Thank you first for giving me some good suggestions on 
> thick-section question I posted last time. Now I met another 
> problem when I did CD4 and FOXP3 dual staining using murine fresh 
> frozen spleen. If I did single staining, both antibodies worked 
> very well. However if I did dual staining, I could only get good 
> result from the first antibody. The second one did not work at 
> all(I mean no specific staining). I used CD4 from BD(rat 
> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. 
> I think there might be something wrong with my serum block(I used 
> normal rat serum block) before I added secondary antibody. Or any 
> other serum block I need to add to decrease the non-specific 
> binding which I have not done yet. Does anyone can give me some 
> suggestions according to my protocol below? How can I get 
> specific staining of the secondary (primary)antibody?  Thank you,
> 
>   Below is the simple protocol I used for dual staining:
>   1. 2% normal goat serum block 20 min
>   2. 1* antibody CD4 1:750 in diluent O/N 4C
>   3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT
>   4. Serum block:  5% normal rat serum 30 min
>   5. 1* antibody FOXP3 1:100 in diluent 1h RT
>   6. Secondary AF488 Donkey anti-rat in TBS 1h RT
> 
>   Use 1xTBS as wash buffer. Before staining, fix tissue in -20C 
> Aceton for 5 min, then airdry.
> 
> 
> Ann Dongtao Fu MD Ph.D
> Lab Manager
> Molecular Pathology core
> Dept. of Pathology
> University of Florida
> 1600 SW Archer Road
> Gainesville, FL 32610
> Lab Phone: 352-273-7752
> FAX: 352-273-7753
> Rm: D11-50
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 

Ann Dongtao Fu MD, Ph.D
Lab Manager
Dept. of Pathology
Lab phone: 352-273-7752
Lab FAX: 352-273-7755
Lab address: D11-50
PO Box: 100275
1600 SW Archer Road
University of Flodrida
Gainesville, FL 32610

_______________________________________________
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"EMF <nvcancer.org>" made the following annotations.
----------------------------------------------------------------------------
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CONFIDENTIALITY NOTICE: This e-mail message, including any
attachments, is for the sole use of the intended 
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and/or privileged information protected by law. If you are 
not the intended recipient, you may not use, copy, or 
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believe you have received this e-mail message in error, 
please contact the sender by reply e-mail and destroy all 
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------------------------------

Message: 19
Date: Thu, 8 Nov 2007 09:47:54 -0800
From: "Tarango, Mark" <mtarango <@t> nvcancer.org>
Subject: RE: [Histonet] B5 substitute
To: "Joanne Mauger" <mauger <@t> email.chop.edu>, gvdobbin <@t> ihis.org,
	Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<5AEC610C1CE02945BD63A395BA763EDE011B6F72 <@t> NVCIEXCH02.NVCI.org>
Content-Type: text/plain; charset=us-ascii

Why not just use 10% NBF?  It's great if you allow enough time for
fixation.  Make sure your bone marrows are fixed well BEFORE
decalcifying and you should have no problems.

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Joanne
Mauger
Sent: Thursday, November 08, 2007 8:51 AM
To: gvdobbin <@t> ihis.org; Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] B5 substitute

Hi ,

What are you using to replace B5 fixative with mercury? I want the best
for hematopoetic tissues, and for immunostains.

Thanks,

Jo Mauger

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

"EMF <nvcancer.org>" made the following annotations.
----------------------------------------------------------------------------
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attachments, is for the sole use of the intended 
recipient(s) and may contain confidential, proprietary, 
and/or privileged information protected by law. If you are 
not the intended recipient, you may not use, copy, or 
distribute this e-mail message or its attachments. If you 
believe you have received this e-mail message in error, 
please contact the sender by reply e-mail and destroy all 
copies of the original message
============================================================================
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------------------------------

Message: 20
Date: Thu, 8 Nov 2007 09:58:22 -0800 (PST)
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] HT position wanted
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <621094.56475.qm <@t> web38209.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

I'm looking for an HT position in the Madison, WI to Rockford, Ill area.
Contract in Milwaukee, WI.

  Thanks,

  Steve

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Message: 21
Date: Thu, 8 Nov 2007 12:59:10 -0500 (EST)
From: "FU,DONGTAO" <fudo <@t> ufl.edu>
Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1211976397.304891194544750373.JavaMail.osg <@t> osgjas03.cns.ufl.edu>
Content-Type: text/plain; format=flowed; charset=us-ascii

Hi, Mark,

  Yes, I am looking at T-regulatory cells of mouse. For CD4, I 
used AF594(red color), for FOXP3 I used AF488(green color). The 
problem for me is for the second primary antibody(or maybe I 
should say for the secondary color) of dual staining, it never 
worked. I do not know which serum I should use to block the 
non-specific staining of the secondary antibody. If you think my 
protocol is imcomplete, could give me some suggestions? Many 
thanks,

Ann

On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" 
<mtarango <@t> nvcancer.org> wrote:

> Looking at T-regulatory cells, huh?  Are you trying to stain both 
> in the
> same color?  I know Foxp3 is nuclear and CD4 is on the cell 
> membrane,
> but two colors might be better.  Your protocol looks incomplete.  
> Mark Adam Tarango HT(ASCP)
> Histology & IHC Supervisor
> Nevada Cancer Institute
> One Breakthrough Way
> Las Vegas, NV  89135
> Direct Line (702) 822-5112
> Treo (702) 759-9229
> Fax (702) 939-7663
>   -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
> FU,DONGTAO
> Sent: Thursday, November 08, 2007 9:13 AM
> To: Histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3
> 
> Hello,
> 
>   Can anyone give me some suggestions on my case? I know a lot of 
> people here have a lot of dual staining experiences. I did dual 
> staining on 2 antibodies from different species in the past. They 
> worked well. But using 2 antibodies from same species is my first 
> time try and I met a big problem. Any suggestions I will be very 
> appreciate.
> 
> Ann
> 
> 
> On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" <fudo <@t> ufl.edu> 
> wrote:
> 
>> Hi, all
>> 
>>   Thank you first for giving me some good suggestions on 
>> thick-section question I posted last time. Now I met another 
>> problem when I did CD4 and FOXP3 dual staining using murine 
>> fresh frozen spleen. If I did single staining, both antibodies 
>> worked very well. However if I did dual staining, I could only 
>> get good result from the first antibody. The second one did not 
>> work at all(I mean no specific staining). I used CD4 from BD(rat 
>> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. 
>> I think there might be something wrong with my serum block(I 
>> used normal rat serum block) before I added secondary antibody. 
>> Or any other serum block I need to add to decrease the 
>> non-specific binding which I have not done yet. Does anyone can 
>> give me some suggestions according to my protocol below? How can 
>> I get specific staining of the secondary (primary)antibody?  
>> Thank you,
>> 
>>   Below is the simple protocol I used for dual staining:
>>   1. 2% normal goat serum block 20 min
>>   2. 1* antibody CD4 1:750 in diluent O/N 4C
>>   3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT
>>   4. Serum block:  5% normal rat serum 30 min
>>   5. 1* antibody FOXP3 1:100 in diluent 1h RT
>>   6. Secondary AF488 Donkey anti-rat in TBS 1h RT
>> 
>>   Use 1xTBS as wash buffer. Before staining, fix tissue in -20C 
>> Aceton for 5 min, then airdry.
>> 
>> 
>> Ann Dongtao Fu MD Ph.D
>> Lab Manager
>> Molecular Pathology core
>> Dept. of Pathology
>> University of Florida
>> 1600 SW Archer Road
>> Gainesville, FL 32610
>> Lab Phone: 352-273-7752
>> FAX: 352-273-7753
>> Rm: D11-50
>> 
>> 
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>> 
>> 
> 
> 
> 
> Ann Dongtao Fu MD, Ph.D
> Lab Manager
> Dept. of Pathology
> Lab phone: 352-273-7752
> Lab FAX: 352-273-7755
> Lab address: D11-50
> PO Box: 100275
> 1600 SW Archer Road
> University of Flodrida
> Gainesville, FL 32610
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> "EMF <nvcancer.org>" made the following annotations.
>
----------------------------------------------------------------------------
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> CONFIDENTIALITY NOTICE: This e-mail message, including any
> attachments, is for the sole use of the intended recipient(s) and 
> may contain confidential, proprietary, and/or privileged 
> information protected by law. If you are not the intended 
> recipient, you may not use, copy, or distribute this e-mail 
> message or its attachments. If you believe you have received this 
> e-mail message in error, please contact the sender by reply 
> e-mail and destroy all copies of the original message
>
============================================================================
==
> 
> 
> 

Ann Dongtao Fu MD, Ph.D
Lab Manager
Dept. of Pathology
Lab phone: 352-273-7752
Lab FAX: 352-273-7755
Lab address: D11-50
PO Box: 100275
1600 SW Archer Road
University of Flodrida
Gainesville, FL 32610

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

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End of Histonet Digest, Vol 48, Issue 11
****************************************