[Histonet] Tracheal wholmount staining

Fri Sep 22 09:00:20 CDT 2006

Dear All

  I am currently screening DKK1 transgenic mice for their effect on tracheal vasculature using the whole mount immunoflouresence.

  I am using Cd31 antibody for that and everything is going well when i do it with retinal whole mount staining but when it comes to trachea, i cant see any blood vessels. What i am thinking is that i am not getting enough penetration.

  Is there anyone who could kindly guide me a complete protocol for whole mount immunoflouresence of trachea.

  Thanks

  Dr.Sohail Ejaz

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Today's Topics:

1. Re: wax temperatures (Bryan Llewellyn)
2. RE: honey as a formalin substitute (Rene J Buesa)
3. RE: Re: [Histonet] Histonet down??? (Zajic Kari)
4. universal negative control for IHC (Susan Bryant)
5. Student writing summary on honey as a fixative and other
comments (Gayle Callis)
6. RE: honey as a formalin substitute (Truscott, Tom)
7. Job Opportunity (Jen, Shirley)
8. Fink-Heimer Silver Stain (Linresearch <@t> aol.com)
9. Re: mounting fluorescent tissue (Markus F. Meyenhofer)
10. RE: honey as a formalin substitute (Edwards, R.E.)
11. RE: FW: Good microtomes replacing JB-4 (Alan Bright)
12. RE: honey as a formalin substitute
(Stephen.Eyres <@t> sanofi-aventis.com)
13. RE: honey as a formalin substitute
(Stephen.Eyres <@t> sanofi-aventis.com)
14. RE: honey as a formalin substitute
(Bartlett, Jeanine (CDC/CCID/NCID))
15. RE: honey as a formalin substitute
(Stephen.Eyres <@t> sanofi-aventis.com)
16. RE: honey as a formalin substitute (Edwards, R.E.)

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

Message: 1
Date: Thu, 21 Sep 2006 09:56:09 -0700
From: Bryan Llewellyn 
Subject: Re: [Histonet] wax temperatures
To: Histonet 
Message-ID: <003401c6dd9e$d68f61d0$130e4246 <@t> yourlk4rlmsu>
Content-Type: text/plain; format=flowed; charset=iso-8859-1;
reply-type=original

Based on my experience with similar waxes, but not Paraplast Xtra itself, 
not much. It gets a little less viscous and likely infiltrates fractionally 
faster, but for usual periods of time involved in processing it has no other 
effect. If kept for too long (weeks or months) the colour changes as a 
result of chemical alteration from the heat. I used to keep some wax of 
this kind in an oven set at about 70C for remelting blocks and such, and I 
experienced no difficulties with it, although it is not a recommended 
practice.

Bryan Llewellyn

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Cote, Laurie
Sent: Thu 9/21/2006 6:54 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] wax temperatures

Does anyone know what happens to Paraplast Xtra if it is 10 degrees
above its melting point?

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Message: 2
Date: Thu, 21 Sep 2006 10:20:25 -0700 (PDT)
From: Rene J Buesa 
Subject: RE: [Histonet] honey as a formalin substitute
To: "Rittman, Barry R" , "Olszewski,
Dawn" , histonet <@t> lists.utsouthwestern.edu
Message-ID: <20060921172025.7835.qmail <@t> web61223.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Barry R:
I cannot agree more with you. You would not have accepted the paper for publication in the form it was accepted neither would have I.
Yes, the JOH is peer reviewed but it seems that peers reviewing this paper gave more weight to publishing the paper than to assure quality of the paper and by doing so have jeopardized the stature of the JOH as the quality vehicle it should be not to mentione the inconsistency between this article and others dealing with esoteric techniques frequently published and wonderful topic articles on December of each year.
Until I resigned from the Editorial board of the JOH I reviewed some papers, and made recommendations that sometimes were followed and sometimes not.
Somebody "drop the ball" at the JOH with this article.
Again, just my opinion.
René J.

"Rittman, Barry R" wrote:
Rene
I was in process of also commenting on this paper to the Journal but perhaps here is as important.

Rene - I agree with all the points that you have written.
To me the images of their formalin fixed tissue do not represent any acceptable level of fixation anywhere I have worked. 
The major problem that I have is that this journal is peer reviewed is it not? My question then is who reviewed this? Based on the article I personally would not have accepted this in its current form for publication - based on the images alone, let alone the other points. 
I am not familiar with the work of the authors but even if they are new to the field I believe that when any papers are submitted for publication they should receive a critical review. This is in everyone's interest. It encourages authors to look very critically at their submitted work and lets them know how their peers will look at the articles.
People should be given every encouragement to submit papers but should also be prepared for a very critical review. We all have tunnel vision when we write papers and benefit from others reviewing these.
Just my opinion also.
Barry

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, September 21, 2006 11:30 AM
To: Olszewski, Dawn; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] honey as a formalin substitute

Dawn:
The article was published in the Journal of Histotechnology, September 2006 issue (vol.29, No.3, pages 173-9).

I read the article and was not really impressed by it because:
1-the honey used came from a source that would not be readily available to everybody, which means that tests with other types/sources of honey will be necessary;
2-there are no comparative methods, no statistical analysis of the results or documented comparisons with standard procedures, and
3- if to substantiate the results we have to judge by the photomicrographs, they are of a very poor quality and what reflect is poor processing. That poor processing could be due to the processing protocol itself or to the fixation the authors claim to have perfomed with honey.
If that is the case, the procedure does not seem to be very promising. I would have require the authors to rewrite some aspects of the paper before publication.
Just my opinion based in what I read.
René J.

"Olszewski, Dawn" wrote:
Help!!!
We have a student in our lab who has been asked to write a summary of the article "The effectiveness of honey as a substitute for formalin in the histological fixation of tissue". Has anyone read this article ( or know where this article can be found) or know anything about this subject matter? If so, any and all info would be appreciated. Thanks in advance. 

Dawn Olszewski
SGMC 
Valdosta, Ga
dawn.olszewski <@t> sgmc.org

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Message: 3
Date: Thu, 21 Sep 2006 13:56:42 -0400
From: "Zajic Kari" 
Subject: RE: Re: [Histonet] Histonet down???
To: "Rene J Buesa" ,

Message-ID:
<095327C7CDBDF64B9E9728A54799091E015CC69F <@t> ORLEV03.hca.corpad.net>
Content-Type: text/plain; charset="iso-8859-1"

Rene` for Histonet president!! :)

he always has all the answers!!! yay! thanks Rene`! :)

Kari Marie Zajic HTL,MLT
Histology Supervisor
Palms West Hospital

email: Kari.Zajic <@t> HCAHealthcare.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Rene J
Buesa
Sent: Thursday, September 21, 2006 12:35 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Fwd: Re: [Histonet] Histonet down???

Fellow Histonetters:
I though you may want to read this e-mail I received from Linda Margraf, to know the reason behind what kept all of us without our "daily dosis of the addictive Histonet".
Regards
René J.

LINDA MARGRAF 
wrote:
Date: Thu, 21 Sep 2006 08:47:42 -0500
From: "LINDA MARGRAF" 

To: 
Subject: Re: [Histonet] Histonet down???

It turns out the problem is with the university email system. They changes some filters and have been blocking all outgoing email from the server. I was still receiving messages from Histonet so it took a while to realize what the problem was. They are working on it and I hope it will be fixed soon. You are on the list and will start getting mail when the filters are fixed. Thanks for your patience. Linda M
Histonet administrator

>>> Rene J Buesa 09/19/06 9:24 AM >>>
Histonet server:
Myself and several user have not received any e-mails via Histonet for several days. Also the archival pages have not been updated since 9/13/06.
Is Histonet out of service?
Please advise
René J. Buesa

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

Message: 4
Date: Thu, 21 Sep 2006 14:47:42 -0400
From: "Susan Bryant" 
Subject: [Histonet] universal negative control for IHC
To: 
Message-ID: <00dd01c6ddae$8181e720$8d01a8c0 <@t> labpath.com>
Content-Type: text/plain; charset="iso-8859-1"

I buy mine from Biocare. It is as they say, both mouse and rabbit IgG. The mouse IgG contains a spectrum of IgG1, IgG2, IgG3, IgG4 and IgM subclasses. 

Susan Bryant
Knoxville, TN

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

Message: 5
Date: Thu, 21 Sep 2006 14:50:23 -0600
From: Gayle Callis 
Subject: [Histonet] Student writing summary on honey as a fixative and
other comments
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20060921142032.01b49708 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Dawn,

Have your student contact the author of this publication for further 
discussion on this topic. I also suggest your student access some of the 
references cited in the publication - it would be a good exercise for doing 
a reference i.e. literature search to broaden their knowledge base.

Philip.Bryant <@t> bromor-tr.wales.nhs.uk

Although honey may not be an ideal formalin substitute and the honey was 
from two species of bees found in Oman, I was intrigued by the topic both 
how it was used in the paper and historically as a preservative and 
dehydrant in other parts of the world. I found the publication unique 
and rather interesting that something like this can be done, maybe without 
the better results of formalin fixation. I also saw the poster at 2006 NSH 
S/C, where many were just as curious as I was honey as a formalin 
substitute, it was a popular poster to visit.

If people are NOT happy with the publication as it is, a letter to the JOH 
editor would be in order and allow the authors to make further comments on 
what points may have been not addressed as they may not be looking in on 
Histonet to see current commentary or critique of their work.

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
406 994-4303 (FAX)

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

Message: 6
Date: Thu, 21 Sep 2006 15:06:39 -0700
From: "Truscott, Tom" 
Subject: RE: [Histonet] honey as a formalin substitute
To: "Rene J Buesa" , "Olszewski, Dawn"
, 
Message-ID:
<35CF12B690D8CA4E95375A36B4E7B44C06CE09 <@t> cvm36.vetmed.wsu.edu>
Content-Type: text/plain; charset="iso-8859-1"

I would also be curious to know if honey collected from plants high in estrogens would affect any type of estrogen staining after processing tissue fixed in honey. Tom Truscott

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, September 21, 2006 8:30 AM
To: Olszewski, Dawn; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] honey as a formalin substitute

Dawn:
The article was published in the Journal of Histotechnology, September 2006 issue (vol.29, No.3, pages 173-9).

I read the article and was not really impressed by it because:
1-the honey used came from a source that would not be readily available to everybody, which means that tests with other types/sources of honey will be necessary;
2-there are no comparative methods, no statistical analysis of the results or documented comparisons with standard procedures, and
3- if to substantiate the results we have to judge by the photomicrographs, they are of a very poor quality and what reflect is poor processing. That poor processing could be due to the processing protocol itself or to the fixation the authors claim to have perfomed with honey.
If that is the case, the procedure does not seem to be very promising. I would have require the authors to rewrite some aspects of the paper before publication.
Just my opinion based in what I read.
René J.

"Olszewski, Dawn" wrote:
Help!!!
We have a student in our lab who has been asked to write a summary of the article "The effectiveness of honey as a substitute for formalin in the histological fixation of tissue". Has anyone read this article ( or know where this article can be found) or know anything about this subject matter? If so, any and all info would be appreciated. Thanks in advance. 

Dawn Olszewski
SGMC 
Valdosta, Ga
dawn.olszewski <@t> sgmc.org

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

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

Message: 7
Date: Thu, 21 Sep 2006 17:54:45 -0700
From: "Jen, Shirley" 
Subject: [Histonet] Job Opportunity
To: 
Message-ID:
<6A39BF27EAB27743887E0425BD51196B0499AB16 <@t> rplmsem1.nala.roche.com>
Content-Type: text/plain; charset="iso-8859-1"

We have a full time histology position available at Roche Palo Alto in California. The position requires working with a variety of laboratory animal tissues for pre-clincal drug safety studies, performing wet tissue trimming to staining for routine paraffin processing, special stains, frozen sections, IHC and other specialized techniques. Interested person please send resume to shirley.jen <@t> roche.com.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Thursday, September 21, 2006 10:10 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 34, Issue 25

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
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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. Fwd: Re: [Histonet] Histonet down??? (Rene J Buesa)
2. Fwd: Re: [Histonet] honey as a formalin substitute (Rene J Buesa)
3. Fwd: Re: [Histonet] Histonet down??? (Rene J Buesa)
4. RE: honey as a formalin substitute (Rittman, Barry R)
5. Re: Histonet Digest, Vol 34, Issue 24 (Steven2146 <@t> aol.com)
6. Re: Tunel staining with acid decalcified samples (Johnson, Teri)
7. RE: Technovit 9100 Kit (Jacqueline D. Garfield)

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

Message: 1
Date: Thu, 21 Sep 2006 09:35:28 -0700 (PDT)
From: Rene J Buesa 
Subject: Fwd: Re: [Histonet] Histonet down???
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20060921163529.68713.qmail <@t> web61220.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Fellow Histonetters:
I though you may want to read this e-mail I received from Linda Margraf, to know the reason behind what kept all of us without our "daily dosis of the addictive Histonet".
Regards
René J.

LINDA MARGRAF 
wrote:
Date: Thu, 21 Sep 2006 08:47:42 -0500
From: "LINDA MARGRAF" 

To: 
Subject: Re: [Histonet] Histonet down???

It turns out the problem is with the university email system. They changes some filters and have been blocking all outgoing email from the server. I was still receiving messages from Histonet so it took a while to realize what the problem was. They are working on it and I hope it will be fixed soon. You are on the list and will start getting mail when the filters are fixed. Thanks for your patience. Linda M Histonet administrator

>>> Rene J Buesa 09/19/06 9:24 AM >>>
Histonet server:
Myself and several user have not received any e-mails via Histonet for several days. Also the archival pages have not been updated since 9/13/06. Is Histonet out of service? Please advise René J. Buesa

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

Message: 2
Date: Thu, 21 Sep 2006 09:46:47 -0700 (PDT)
From: Rene J Buesa 
Subject: Fwd: Re: [Histonet] honey as a formalin substitute
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20060921164647.42194.qmail <@t> web61217.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Rene J Buesa wrote: Date: Thu, 21 Sep 2006 09:29:55 -0700 (PDT)
From: Rene J Buesa 
Subject: Re: [Histonet] honey as a formalin substitute
To: "Olszewski, Dawn" , 
histonet <@t> lists.utsouthwestern.edu

Dawn:
The article was published in the Journal of Histotechnology, September 2006 issue (vol.29, No.3, pages 173-9).

I read the article and was not really impressed by it because:
1-the honey used came from a source that would not be readily available to everybody, which means that tests with other types/sources of honey will be necessary;
2-there are no comparative methods, no statistical analysis of the results or documented comparisons with standard procedures, and
3- if to substantiate the results we have to judge by the photomicrographs, they are of a very poor quality and what reflect is poor processing. That poor processing could be due to the processing protocol itself or to the fixation the authors claim to have perfomed with honey.

=== message truncated ===

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