[Histonet] RE: Histonet Digest, Vol 63, Issue 24

Collins, Michael mike.collins <@t> imperial.ac.uk
Mon Feb 16 10:18:58 CST 2009


Hi Paul!
In the 60s I used to make the Barger and de Lameter Schiffs using thionyl chloride to produce sulphurous acid. I used activated charcoal and filtered with a Buchner funnel connected to a fast flowing cold tap. Vivid staining!
Mike CollinS.

-----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: 14 February 2009 17:55
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 63, Issue 24

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Today's Topics:

   1. hand processing schedule late mouse embryos (Nicole Collette)
   2. CD31 on FFPE mouse tissue (Kim Merriam)
   3. Re: CD31 on FFPE mouse tissue (Jackie M O'Connor)
   4. need controls (Steven Coakley)
   5. RE: CD31 on FFPE mouse tissue (Patsy Ruegg)
   6. Histology in tough economy (O'Donnell, Bill)
   7. Re: Histology in tough economy (Peter Carroll)
   8. Elisa testing (Santiago, Albert)
   9. Storage of Tissues (Breeden, Sara)
  10. Re: CD31 on FFPE mouse tissue (Colleen Forster)
  11. Re: CD31 on FFPE mouse tissue (Andrea Hooper)
  12. Re: Histology in tough economy (Geoff McAuliffe)
  13. AUTO: Jacquelyn Grewe/Staff/OhioHealth is out of the	office .
      (JGREWE <@t> OhioHealth.com)
  14. RE: Histology in tough economy (O'Donnell, Bill)
  15. Re: CD31 on FFPE mouse tissue (nancy lowen)
  16. Re: need controls (Jaime Plata)
  17. freezing paraffin blocks (Jennifer Anderson)
  18. Is there anyone out there that is using the VIP6 from	Sakura?
      (Bull, Jennifer L.)
  19. Fwd: Can you place on the histonet? (LINDA MARGRAF)
  20. fume exposure (zodiac29 <@t> comcast.net)
  21. RE: fume exposure (Smith, Allen)
  22. Re: hand processing schedule late mouse embryos (Rene J Buesa)
  23. Re: need controls (Rene J Buesa)
  24. Re: freezing paraffin blocks (Rene J Buesa)
  25. Problem with Schiff's reagent (pbrunko <@t> siue.edu)


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

Message: 1
Date: Fri, 13 Feb 2009 10:25:47 -0800
From: Nicole Collette <collette2 <@t> mail.llnl.gov>
Subject: [Histonet] hand processing schedule late mouse embryos
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <p0623091bc5bb5e5b0df5@[128.115.150.224]>
Content-Type: text/plain; charset="us-ascii" ; format="flowed"

HI, All,

One project finished, another just beginning. I am about to embark on 
a journey into the land of immunohistochemistry, with late mouse 
embryos E14.5, E16.5, P0 to examine bone markers in conjunction with 
LacZ and/or GFP.

We have sadly lost our cryostat (so IHC for the GFP on paraffin 
sections), and our tissue processor - both belonged to a friendly 
investigator down the hall who has moved on. So, I am processing by 
hand.

For hand-processing, I have had to do some rigging, and I do the wax 
steps in a hyb oven to try to keep the wax (TissuePrep, Fisher) at 
around 63-65C, while trying my best to keep the molds, cassettes, and 
tools with as few giant globs of solidifying wax as possible. As a 
result of using the hyb oven, we are forced to use Clearene 
(D-limonene), --or some other xylene substitute that could be 
recommended--,  instead of xylene for the processing. If anyone has a 
recommendation for a better alternative there (aside from a tissue 
processor which will have to wait at least until the next grant gets 
funded--oooh, unless someone has an old one they want to donate, 
preferably table-top), I'm all ears.

My schedule was given to me by a friend who does cartilage, no older 
than E14.5, and are basically half hour steps for each ethanol, half 
hour steps for 3 wax steps at the end. Will this be enough time for 
infiltration of older samples without vacuum? Should I increase my 
steps to 1 hour for these older embryos? I am optimizing my fixation 
at 1hour/mm thickness, with the embryos skinned (4% paraformaldehyde 
in PBS, I decided to start here since I don't yet know much about the 
problems I might encounter with a particular antigen). I have tried 
the 30 minute schedule with adult decalcified bones and have not had 
fantastic sections. I suspect it could be incomplete washing of the 
EDTA before infiltration, but it's possible that the processing 
schedule is just not long enough. Any advice?

Thanks in advance! Happy Friday!

Sincerely,
Nicole Collette
Lawrence Livermore National Lab/ UC Berkeley



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

Message: 2
Date: Fri, 13 Feb 2009 10:27:16 -0800 (PST)
From: Kim Merriam <kmerriam2003 <@t> yahoo.com>
Subject: [Histonet] CD31 on FFPE mouse tissue
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <799092.72818.qm <@t> web50304.mail.re2.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

What is everyone doing for CD31 on FFPE mouse tissue?  We are looking to do IF, but a DAB protocol would be just as good.
 
Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA


      

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

Message: 3
Date: Fri, 13 Feb 2009 12:33:15 -0600
From: Jackie M O'Connor <Jackie.O'Connor <@t> abbott.com>
Subject: Re: [Histonet] CD31 on FFPE mouse tissue
To: kmerriam2003 <@t> yahoo.com
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>,
	histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID:
	<OF5AEA4DD5.2E104C64-ON8625755C.0065DDE5-8625755C.0065F2C4 <@t> abbott.com>
Content-Type: text/plain; charset="US-ASCII"

Biocare Medical is supposed to have a good CD31 for FFPE mouse.   I don't 
have proof of this, but I would contact Biocare.





Kim Merriam <kmerriam2003 <@t> yahoo.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
02/13/2009 12:27 PM
Please respond to
kmerriam2003 <@t> yahoo.com


To
Histonet <histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] CD31 on FFPE mouse tissue






What is everyone doing for CD31 on FFPE mouse tissue?  We are looking to 
do IF, but a DAB protocol would be just as good.
 
Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA


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



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

Message: 4
Date: Fri, 13 Feb 2009 10:51:01 -0800 (PST)
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <795858.41301.qm <@t> web38201.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

I work in a DermPath lab.  Currently we purchase all our controls.  Some are ok but some, especially the grams are not.  We'd like to start sectioning our own but all the specimens we get are skins.


      

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

Message: 5
Date: Fri, 13 Feb 2009 11:54:05 -0700
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: [Histonet] CD31 on FFPE mouse tissue
To: "'Jackie M O'Connor'" <Jackie.O'Connor <@t> abbott.com>,
	<kmerriam2003 <@t> yahoo.com>
Cc: 'Histonet' <histonet <@t> lists.utsouthwestern.edu>,
	histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID: <BFD4F8E2F97B4110B443293E0F417DD0 <@t> prueggihctechlt>
Content-Type: text/plain;	charset="us-ascii"

I got a sample of the BC cd31 for mouse but have not had a chance to try it
yet, I will let you know.  I have heard from others on the IHC Resource
Group list server that it is pretty good but does miss some of the very
early vessels forming with implants.
NSH members can join the IHCRG online at www.ihcrg.org 

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jackie M
O'Connor
Sent: Friday, February 13, 2009 11:33 AM
To: kmerriam2003 <@t> yahoo.com
Cc: Histonet; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] CD31 on FFPE mouse tissue

Biocare Medical is supposed to have a good CD31 for FFPE mouse.   I don't 
have proof of this, but I would contact Biocare.





Kim Merriam <kmerriam2003 <@t> yahoo.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
02/13/2009 12:27 PM
Please respond to
kmerriam2003 <@t> yahoo.com


To
Histonet <histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] CD31 on FFPE mouse tissue






What is everyone doing for CD31 on FFPE mouse tissue?  We are looking to 
do IF, but a DAB protocol would be just as good.
 
Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA


 
_______________________________________________
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




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

Message: 6
Date: Fri, 13 Feb 2009 11:56:20 -0700
From: "O'Donnell, Bill" <billodonnell <@t> catholichealth.net>
Subject: [Histonet] Histology in tough economy
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<A6871E977189984E9893DAC85D2C160E675AAF <@t> chimsx04.CHI.catholichealth.net>
	
Content-Type: text/plain; charset=us-ascii

Greetings fellow hsitology professionals. 

In today's economic climate, many of us are finding ourselves faced with
changes in our policies, staffing, spending, etc. 

To that end I have started a histology blog.

This is a place to see what is happening in other labs and to share your
own comments. 

This blog is meant to be TEMPORARY in nature. IT IS NOT MEANT TO REPLACE
THE HISTONET, as it is and remains a fine vehicle for communication and
has a long history of serving our community world-wide.

It is a place where threads can be long, unfettered and anonymous.

Allowing anonimity will free us to feel comfortable in our expressions.

People will need to register in order to comment.  However, you choose
your username and password. A real e-mail address is also needed, but if
it is generic,, that is, does not contaiin your name or institution,
that is fine. (example: tech <@t> hotmail.com) 

The e-mail will only be used to contact you concerning this site and to
send you data once it has been compiled. It will not be sold or used iin
any other fashion.

All registered users will be allowed contributer status.

We hope to hear from clinical, research, and veterinary histologists,
pathologists or administrators.

Sales and staffing professionals are also most welcome.

Every 30 days or so the accumulated data will be shared with each
subscriber in the hopes that our profession might benefit from our
collective wisdom and experiences.

I am not collecting this info for any personal gain, nor is there any
other motive than that of helping out our profession.

The link is http://histoblog.deaconbill.com/

I will resend this message from time-to-time so that all might have a
chance to participate.

If you have any questions regarding the use of this blog, please e-mail
me directly at bill <@t> deaconbill.com





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

Message: 7
Date: Fri, 13 Feb 2009 14:05:48 -0500
From: Peter Carroll <carrolpb <@t> umdnj.edu>
Subject: Re: [Histonet] Histology in tough economy
To: "O'Donnell, Bill" <billodonnell <@t> catholichealth.net>,	Histonet
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4995C48C.7070602 <@t> umdnj.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

here's hoping you spell-check your blog, bill ;)

O'Donnell, Bill wrote:
> Greetings fellow hsitology professionals. 
>
> In today's economic climate, many of us are finding ourselves faced with
> changes in our policies, staffing, spending, etc. 
>
> To that end I have started a histology blog.
>
> This is a place to see what is happening in other labs and to share your
> own comments. 
>
> This blog is meant to be TEMPORARY in nature. IT IS NOT MEANT TO REPLACE
> THE HISTONET, as it is and remains a fine vehicle for communication and
> has a long history of serving our community world-wide.
>
> It is a place where threads can be long, unfettered and anonymous.
>
> Allowing anonimity will free us to feel comfortable in our expressions.
>
> People will need to register in order to comment.  However, you choose
> your username and password. A real e-mail address is also needed, but if
> it is generic,, that is, does not contaiin your name or institution,
> that is fine. (example: tech <@t> hotmail.com) 
>
> The e-mail will only be used to contact you concerning this site and to
> send you data once it has been compiled. It will not be sold or used iin
> any other fashion.
>
> All registered users will be allowed contributer status.
>
> We hope to hear from clinical, research, and veterinary histologists,
> pathologists or administrators.
>
> Sales and staffing professionals are also most welcome.
>
> Every 30 days or so the accumulated data will be shared with each
> subscriber in the hopes that our profession might benefit from our
> collective wisdom and experiences.
>
> I am not collecting this info for any personal gain, nor is there any
> other motive than that of helping out our profession.
>
> The link is http://histoblog.deaconbill.com/
>
> I will resend this message from time-to-time so that all might have a
> chance to participate.
>
> If you have any questions regarding the use of this blog, please e-mail
> me directly at bill <@t> deaconbill.com
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>   




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

Message: 8
Date: Fri, 13 Feb 2009 14:11:27 -0500
From: "Santiago, Albert" <Albert.Santiago <@t> uphs.upenn.edu>
Subject: [Histonet] Elisa testing
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<BB0F4D2041EED9458AD95A538DBC4CAE02FA73F6 <@t> uphsmbx6.UPHS.PENNHEALTH.PRV>
	
Content-Type: text/plain;	charset="us-ascii"

Hello all and Happy Valentine's Day, We're in the process of starting to
do Elisa testing and I my question to anyone out there who is already
doing it, did you go through anykind of proficiency testing and/or
compliance preparedness for JCAHO or CAP before beginning to do Elisas?
Thanks for your help....

 

Albert Santiago, HT (ASCP)

Laboratory Supervisor

Dermatopathology

Phone 215-662-6539

Fax     215-662-6150

 

 

 



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

Message: 9
Date: Fri, 13 Feb 2009 12:18:15 -0700
From: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>
Subject: [Histonet] Storage of Tissues
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4D14F0FC9316DD41972D5F03C070908B017E66C5 <@t> nmdamailsvr.nmda.ad.nmsu.edu>
	
Content-Type: text/plain;	charset="US-ASCII"

We researched the subject and this is what we do: 10 years for slides, 7
years for blocks.  Wet tissues ("surgicals", "biopsies") are kept for
about six weeks after final report is issued. Specimens are boxed daily
and marked with date; the lead tech in Receiving checks one or two
specimens from each box after a couple weeks to make sure final report
has been issued, then marks them with disposal date.  Necropsy tissues
are marked with a disposal date of six weeks from the date I get the
slides back from the pathologist(s).  Slides are sealed into cardboard
boxes and marked "Glass Waste" and disposed of via the City's solid
waste department; blocks are sealed into cardboard boxes, marked
"Paraffin Waste" and also disposed of via the City's solid waste dept.
Hope this helps.

Sally Breeden, HT(ASCP)
NM Dept. of Agriculture
Veterinary Diagnostic Services
PO Box 4700
Albuquerque, NM  87106
505-841-2576




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

Message: 10
Date: Fri, 13 Feb 2009 13:49:34 -0600
From: Colleen Forster <cforster <@t> umn.edu>
Subject: Re: [Histonet] CD31 on FFPE mouse tissue
To: Jackie M O'Connor <Jackie.O'Connor <@t> abbott.com>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>,
	histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID: <4995CECE.4020504 <@t> umn.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Biocare does have a very good antibody and kit for this and I can tell 
you it works very nicely if you use their kit and primary.

Colleen Forster
612-626-1930
U of MN


Jackie M O'Connor wrote:
> Biocare Medical is supposed to have a good CD31 for FFPE mouse.   I don't 
> have proof of this, but I would contact Biocare.
>
>
>
>
>
> Kim Merriam <kmerriam2003 <@t> yahoo.com> 
> Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
> 02/13/2009 12:27 PM
> Please respond to
> kmerriam2003 <@t> yahoo.com
>
>
> To
> Histonet <histonet <@t> lists.utsouthwestern.edu>
> cc
>
> Subject
> [Histonet] CD31 on FFPE mouse tissue
>
>
>
>
>
>
> What is everyone doing for CD31 on FFPE mouse tissue?  We are looking to 
> do IF, but a DAB protocol would be just as good.
>  
> Thanks,
> Kim
>
> Kim Merriam, MA, HT(ASCP)QIHC
> Cambridge, MA
>
>
>  
> _______________________________________________
> 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
>
>   




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

Message: 11
Date: Fri, 13 Feb 2009 15:00:12 -0500
From: "Andrea Hooper" <anh2006 <@t> med.cornell.edu>
Subject: Re: [Histonet] CD31 on FFPE mouse tissue
To: Histonet <histonet <@t> lists.utsouthwestern.edu>,	Kim Merriam
	<kmerriam2003 <@t> yahoo.com>
Message-ID: <p06200708c5bb81598a29@[140.251.51.13]>
Content-Type: text/plain; charset=us-ascii; format=flowed

Does it have to be CD31? I have a bunch of good protocols for mouse 
vessels but each tissue requires a different antibody. CD31 in FFPE 
is never reliable in my experience. Which tissue are you using?

Andrea


>>
>>
>>Kim Merriam <kmerriam2003 <@t> yahoo.com> Sent by: 
>>histonet-bounces <@t> lists.utsouthwestern.edu
>>02/13/2009 12:27 PM
>>Please respond to
>>kmerriam2003 <@t> yahoo.com
>>
>>
>>
>>What is everyone doing for CD31 on FFPE mouse tissue?  We are 
>>looking to do IF, but a DAB protocol would be just as good.
>>  Thanks,
>>Kim
>>
>>Kim Merriam, MA, HT(ASCP)QIHC
>>Cambridge, MA

-- 



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

Message: 12
Date: Fri, 13 Feb 2009 15:25:15 -0500
From: Geoff McAuliffe <mcauliff <@t> umdnj.edu>
Subject: Re: [Histonet] Histology in tough economy
To: Peter Carroll <carrolpb <@t> umdnj.edu>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4995D72B.1020407 <@t> umdnj.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Here's hoping you don't have too many nit pickers criticizing your blog.

"It's a mighty poor speller who can't think of more than one way to 
spell a word."
Warren G. Harding, President of the US, 1920-1923.

Peter Carroll wrote:
> here's hoping you spell-check your blog, bill ;)
>
> O'Donnell, Bill wrote:
>> Greetings fellow hsitology professionals.
>> In today's economic climate, many of us are finding ourselves faced with
>> changes in our policies, staffing, spending, etc.
>> To that end I have started a histology blog.
>>
>> This is a place to see what is happening in other labs and to share your
>> own comments.
>> This blog is meant to be TEMPORARY in nature. IT IS NOT MEANT TO REPLACE
>> THE HISTONET, as it is and remains a fine vehicle for communication and
>> has a long history of serving our community world-wide.
>>
>> It is a place where threads can be long, unfettered and anonymous.
>>
>> Allowing anonimity will free us to feel comfortable in our expressions.
>>
>> People will need to register in order to comment.  However, you choose
>> your username and password. A real e-mail address is also needed, but if
>> it is generic,, that is, does not contaiin your name or institution,
>> that is fine. (example: tech <@t> hotmail.com)
>> The e-mail will only be used to contact you concerning this site and to
>> send you data once it has been compiled. It will not be sold or used iin
>> any other fashion.
>>
>> All registered users will be allowed contributer status.
>>
>> We hope to hear from clinical, research, and veterinary histologists,
>> pathologists or administrators.
>>
>> Sales and staffing professionals are also most welcome.
>>
>> Every 30 days or so the accumulated data will be shared with each
>> subscriber in the hopes that our profession might benefit from our
>> collective wisdom and experiences.
>>
>> I am not collecting this info for any personal gain, nor is there any
>> other motive than that of helping out our profession.
>>
>> The link is http://histoblog.deaconbill.com/
>>
>> I will resend this message from time-to-time so that all might have a
>> chance to participate.
>>
>> If you have any questions regarding the use of this blog, please e-mail
>> me directly at bill <@t> deaconbill.com
>>
>>
>>
>> _______________________________________________
>> 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
>
>


-- 
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583 
mcauliff <@t> umdnj.edu
**********************************************





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

Message: 13
Date: Fri, 13 Feb 2009 16:00:23 -0500
From: JGREWE <@t> OhioHealth.com
Subject: [Histonet] AUTO: Jacquelyn Grewe/Staff/OhioHealth is out of
	the	office . 
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<OF7F46B243.13870252-ON8525755C.0073648F-8525755C.0073648E <@t> ohiohealth.com>
	
Content-Type: text/plain; charset=US-ASCII


I will be out of the office starting  02/11/2009 and will not return until
02/16/2009.

I will respond to your message when I return.


Note: This is an automated response to your message  "Histonet Digest, Vol
63, Issue 23" sent on 2/13/2009 1:08:10 PM.

You will receive a notification for each message you send to this person
while the person is away.




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

Message: 14
Date: Fri, 13 Feb 2009 14:27:33 -0700
From: "O'Donnell, Bill" <billodonnell <@t> catholichealth.net>
Subject: RE: [Histonet] Histology in tough economy
To: "Geoff McAuliffe" <mcauliff <@t> umdnj.edu>,	"Peter Carroll"
	<carrolpb <@t> umdnj.edu>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<A6871E977189984E9893DAC85D2C160E675AD9 <@t> chimsx04.CHI.catholichealth.net>
	
Content-Type: text/plain; charset=us-ascii

I need spelling nit-pickers, othewise I look really stoopid! Have a
great weekend, and don't forget to sign up for the blog.
Bill 

-----Original Message-----
From: Geoff McAuliffe [mailto:mcauliff <@t> umdnj.edu] 
Sent: Friday, February 13, 2009 2:25 PM
To: Peter Carroll
Cc: O'Donnell, Bill; Histonet
Subject: Re: [Histonet] Histology in tough economy

Here's hoping you don't have too many nit pickers criticizing your blog.

"It's a mighty poor speller who can't think of more than one way to
spell a word."
Warren G. Harding, President of the US, 1920-1923.

Peter Carroll wrote:
> here's hoping you spell-check your blog, bill ;)
>
> O'Donnell, Bill wrote:
>> Greetings fellow hsitology professionals.
>> In today's economic climate, many of us are finding ourselves faced 
>> with changes in our policies, staffing, spending, etc.
>> To that end I have started a histology blog.
>>
>> This is a place to see what is happening in other labs and to share 
>> your own comments.
>> This blog is meant to be TEMPORARY in nature. IT IS NOT MEANT TO 
>> REPLACE THE HISTONET, as it is and remains a fine vehicle for 
>> communication and has a long history of serving our community
world-wide.
>>
>> It is a place where threads can be long, unfettered and anonymous.
>>
>> Allowing anonimity will free us to feel comfortable in our
expressions.
>>
>> People will need to register in order to comment.  However, you 
>> choose your username and password. A real e-mail address is also 
>> needed, but if it is generic,, that is, does not contaiin your name 
>> or institution, that is fine. (example: tech <@t> hotmail.com) The e-mail 
>> will only be used to contact you concerning this site and to send you

>> data once it has been compiled. It will not be sold or used iin any 
>> other fashion.
>>
>> All registered users will be allowed contributer status.
>>
>> We hope to hear from clinical, research, and veterinary histologists,

>> pathologists or administrators.
>>
>> Sales and staffing professionals are also most welcome.
>>
>> Every 30 days or so the accumulated data will be shared with each 
>> subscriber in the hopes that our profession might benefit from our 
>> collective wisdom and experiences.
>>
>> I am not collecting this info for any personal gain, nor is there any

>> other motive than that of helping out our profession.
>>
>> The link is http://histoblog.deaconbill.com/
>>
>> I will resend this message from time-to-time so that all might have a

>> chance to participate.
>>
>> If you have any questions regarding the use of this blog, please 
>> e-mail me directly at bill <@t> deaconbill.com
>>
>>
>>
>> _______________________________________________
>> 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
>
>


--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
**********************************************






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

Message: 15
Date: Fri, 13 Feb 2009 13:43:26 -0800 (PST)
From: nancy lowen <claycal44 <@t> yahoo.com>
Subject: Re: [Histonet] CD31 on FFPE mouse tissue
To: kmerriam2003 <@t> yahoo.com, Jackie M O'Connor
	<Jackie.O'Connor <@t> abbott.com>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>,
	histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID: <571567.63945.qm <@t> web65607.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

I have used the antibody and kit from Biocare for CD31 and got good results.  Simple to use.
Nancy.lowen <@t> med.va.gov

--- On Fri, 2/13/09, Jackie M O'Connor <Jackie.O'Connor <@t> abbott.com> wrote:

From: Jackie M O'Connor <Jackie.O'Connor <@t> abbott.com>
Subject: Re: [Histonet] CD31 on FFPE mouse tissue
To: kmerriam2003 <@t> yahoo.com
Cc: "Histonet" <histonet <@t> lists.utsouthwestern.edu>, histonet-bounces <@t> lists.utsouthwestern.edu
Date: Friday, February 13, 2009, 10:33 AM

Biocare Medical is supposed to have a good CD31 for FFPE mouse.   I don't 
have proof of this, but I would contact Biocare.





Kim Merriam <kmerriam2003 <@t> yahoo.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
02/13/2009 12:27 PM
Please respond to
kmerriam2003 <@t> yahoo.com


To
Histonet <histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] CD31 on FFPE mouse tissue






What is everyone doing for CD31 on FFPE mouse tissue?  We are looking to 
do IF, but a DAB protocol would be just as good.
 
Thanks,
Kim

Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA


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

Message: 16
Date: Fri, 13 Feb 2009 14:26:03 -0800 (PST)
From: Jaime Plata <enrriq88 <@t> yahoo.com>
Subject: Re: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu, Steven Coakley
	<sjchtascp <@t> yahoo.com>
Message-ID: <69741.96460.qm <@t> web50410.mail.re2.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

GI (small intestine) biopsies are other possibilities associated to skin as control.
When you mention Grams, are you referring to Bact's positive and negative.? If this is the case you can make them with blood Agar (microbiology) and contaminate the tissue with them. Or use Large intestine ther will be many you jus need to look for them.
 
--- On Fri, 2/13/09, Steven Coakley <sjchtascp <@t> yahoo.com> wrote:
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu
Date: Friday, February 13, 2009, 1:51 PM

I work in a DermPath lab.  Currently we purchase all our controls.  Some are
Ok but some, especially the grams are not.  We'd like to start sectioning.
Our own but all the specimens we get are skins.
 
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


      

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

Message: 17
Date: Fri, 13 Feb 2009 14:43:25 -0800
From: "Jennifer Anderson" <janderson <@t> halozyme.com>
Subject: [Histonet] freezing paraffin blocks
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<A037532CC3E3974CB0AB2420B928DACD01FD89EF <@t> HTIEXCHANGE.hti.com>
Content-Type: text/plain;	charset="us-ascii"

Hi All.

 

I've just started sectioning Bouin's fixed paraffin embedded
human/mouse/rat/pig skin again, tissues processed in a softer wax than
I'm used to, so I've been putting the blocks in the freezer at -20 prior
to sectioning.  Will such cold deleteriously affect the blocks or
tissue?

 

Also, is there any harm in cleaning the water bath with a little xylene
to remove any wax build-up?

 

Thanks a lot.

 

Jennifer M. Anderson, Scientist

Halozyme Therapeutics, Inc.

11404 Sorrento Valley Road

San Diego, CA 92121

858-704-8333

janderson <@t> halozyme.com

 

 


The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed.  Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege.  Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws.  If you received this in error, please contact the sender and delete/destroy this email.


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

Message: 18
Date: Fri, 13 Feb 2009 15:52:51 -0800
From: "Bull, Jennifer L." <Jennifer.Bull <@t> northwestpathology.com>
Subject: [Histonet] Is there anyone out there that is using the VIP6
	from	Sakura?
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<85760CECEC18444BB95F26D5E88DAEAA21FF216F8C <@t> hinet2.hinet.org>
Content-Type: text/plain; charset="us-ascii"

Is there anyone out there that is currently using the VIP6 that could give me feedback on processing times for small biopsoes and surgicals? We are trying to get a realistic comparison of the Perloris to the tried and true from Sakura. Thanks in advance!

Jennifer Bull
jennifer.bull <@t> nwpathology.com



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

Message: 19
Date: Fri, 13 Feb 2009 18:42:06 -0600
From: "LINDA MARGRAF" <LINDA.MARGRAF <@t> childrens.com>
Subject: [Histonet] Fwd: Can you place on the histonet?
To: <histonet <@t> lists.utsouthwestern.edu>
Cc: cbarone <@t> NEMOURS.ORG
Message-ID: <4995BF0D.F783.00DA.0 <@t> childrens.com>
Content-Type: text/plain; charset="us-ascii"

Here's a message Carol wanted posted to the list.  Please respond to her and not me. Thanks
Linda M
Histonet administrator

>>> "Barone, Carol " <cbarone <@t> NEMOURS.ORG> 2/13/2009 10:33 AM >>>

Anyone using "bead" detection systems with readers for tissue histology. I have investigators interested in these for slides? Need infoFAST...for grant submission...Help!!!!
</pre>	<span style="font-weight: bold;">Please consider the environment before printing this e-mail</span><br />
	<br />
	
	<span style="font-size: 8pt;">This e-mail, facsimile, or letter and any files or attachments transmitted with it contains<br />
		information that is confidential and privileged. This information is intended only for the use of the <br />
		individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further <br />

		disclosures are prohibited without proper authorization. If you are not the intended recipient, any <br />
		disclosure, copying, printing, or use of this information is strictly prohibited and possibly a <br />
		violation of federal or state law and regulations. If you have received this information in error, <br />
		please notify Children's Medical Center Dallas immediately at 214-456-4444 or via e-mail at <br />
		privacy <@t> childrens.com. Children's Medical Center Dallas and its affiliates hereby claim all <br />
		applicable privileges related to this information.</span><br />

	<br />
</html>


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

Message: 20
Date: Sat, 14 Feb 2009 11:46:16 +0000 (UTC)
From: zodiac29 <@t> comcast.net
Subject: [Histonet] fume exposure
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<1317950559.2115121234611976799.JavaMail.root <@t> sz0062a.westchester.pa.mail.comcast.net>
	
Content-Type: text/plain; charset=utf-8




To All, 



I work in a lab where I stain and coverslip everything by hand. I coverslip under a fume hood so there isn't much of an issue there. The main issue is my exposue to fumes while staining. I do not stain under a fume hood. My exposure times to these c hemicals is increasing becuase our volume is steadly increasing as well. I can no longer tolerate these fumes and am expirenc ing difficulty breathing and frequent headaches.What are labs that stain there slides  by hand doing (types of masks, hoods) to minimize their exposure. If you can recommed any specifics I would appreciate it. 



Thank you in advance 

Jenny 





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

Message: 21
Date: Sat, 14 Feb 2009 10:26:21 -0500
From: "Smith, Allen" <asmith <@t> mail.barry.edu>
Subject: RE: [Histonet] fume exposure
To: "'zodiac29 <@t> comcast.net'" <zodiac29 <@t> comcast.net>
Cc: "'Histonet <@t> lists.utsouthwestern.edu'"
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<E4132130AC2F764D8C173C5400D530429FA4BD5687 <@t> exchsrv02.barrynet.barry.edu>
	
Content-Type: text/plain; charset="utf-8"

The only staining solutions whose fumes bother me are ammonium sulfide and sulfurous acid.  I put the staining jars of these in the fume hood and carry the staining jar with the slides to the fume hood.  I bring the water rinse jar to the fume hood to take the slides out.  While sulfurous acid is merely irritating, ammonium sulfide is a mitochondrial poison (about 10% as poisonous as prussic acid).
Allen A. Smith
Professor of Anatomy
Barry University School of Podiatric Medicine

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of zodiac29 <@t> comcast.net
Sent: Saturday, February 14, 2009 6:46 AM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] fume exposure




To All,



I work in a lab where I stain and coverslip everything by hand. I coverslip under a fume hood so there isn't much of an issue there. The main issue is my exposue to fumes while staining. I do not stain under a fume hood. My exposure times to these c hemicals is increasing becuase our volume is steadly increasing as well. I can no longer tolerate these fumes and am expirenc ing difficulty breathing and frequent headaches.What are labs that stain there slides  by hand doing (types of masks, hoods) to minimize their exposure. If you can recommed any specifics I would appreciate it.



Thank you in advance

Jenny



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


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

Message: 22
Date: Sat, 14 Feb 2009 08:21:39 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] hand processing schedule late mouse embryos
To: histonet <@t> lists.utsouthwestern.edu,	Nicole Collette
	<collette2 <@t> mail.llnl.gov>
Message-ID: <683350.23959.qm <@t> web65713.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Under separate cover I am sending a protocol that will work well with your subjects.
Since you are doing manual processing you can easily use the reagents I recommend and eliminate xylene and its terpene and alkane based substitutes.
René J.

--- On Fri, 2/13/09, Nicole Collette <collette2 <@t> mail.llnl.gov> wrote:

From: Nicole Collette <collette2 <@t> mail.llnl.gov>
Subject: [Histonet] hand processing schedule late mouse embryos
To: histonet <@t> lists.utsouthwestern.edu
Date: Friday, February 13, 2009, 1:25 PM

HI, All,

One project finished, another just beginning. I am about to embark on a journey
into the land of immunohistochemistry, with late mouse embryos E14.5, E16.5, P0
to examine bone markers in conjunction with LacZ and/or GFP.

We have sadly lost our cryostat (so IHC for the GFP on paraffin sections), and
our tissue processor - both belonged to a friendly investigator down the hall
who has moved on. So, I am processing by hand.

For hand-processing, I have had to do some rigging, and I do the wax steps in a
hyb oven to try to keep the wax (TissuePrep, Fisher) at around 63-65C, while
trying my best to keep the molds, cassettes, and tools with as few giant globs
of solidifying wax as possible. As a result of using the hyb oven, we are forced
to use Clearene (D-limonene), --or some other xylene substitute that could be
recommended--,  instead of xylene for the processing. If anyone has a
recommendation for a better alternative there (aside from a tissue processor
which will have to wait at least until the next grant gets funded--oooh, unless
someone has an old one they want to donate, preferably table-top), I'm all
ears.

My schedule was given to me by a friend who does cartilage, no older than
E14.5, and are basically half hour steps for each ethanol, half hour steps for 3
wax steps at the end. Will this be enough time for infiltration of older samples
without vacuum? Should I increase my steps to 1 hour for these older embryos? I
am optimizing my fixation at 1hour/mm thickness, with the embryos skinned (4%
paraformaldehyde in PBS, I decided to start here since I don't yet know much
about the problems I might encounter with a particular antigen). I have tried
the 30 minute schedule with adult decalcified bones and have not had fantastic
sections. I suspect it could be incomplete washing of the EDTA before
infiltration, but it's possible that the processing schedule is just not
long enough. Any advice?

Thanks in advance! Happy Friday!

Sincerely,
Nicole Collette
Lawrence Livermore National Lab/ UC Berkeley

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

Message: 23
Date: Sat, 14 Feb 2009 08:25:13 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu, Steven Coakley
	<sjchtascp <@t> yahoo.com>,	enrriq88 <@t> yahoo.com
Message-ID: <804168.42750.qm <@t> web65705.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

The same appendix that you can use as an H&E+ control, usually contains Gram +and- bacteria.
René J.

--- On Fri, 2/13/09, Jaime Plata <enrriq88 <@t> yahoo.com> wrote:

From: Jaime Plata <enrriq88 <@t> yahoo.com>
Subject: Re: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu, "Steven Coakley" <sjchtascp <@t> yahoo.com>
Date: Friday, February 13, 2009, 5:26 PM

GI (small intestine) biopsies are other possibilities associated to skin as
control.
When you mention Grams, are you referring to Bact's positive and negative.?
If this is the case you can make them with blood Agar (microbiology) and
contaminate the tissue with them. Or use Large intestine ther will be many you
jus need to look for them.
 
--- On Fri, 2/13/09, Steven Coakley <sjchtascp <@t> yahoo.com> wrote:
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] need controls
To: Histonet <@t> lists.utsouthwestern.edu
Date: Friday, February 13, 2009, 1:51 PM

I work in a DermPath lab.  Currently we purchase all our controls.  Some are
Ok but some, especially the grams are not.  We'd like to start sectioning.
Our own but all the specimens we get are skins.
 
_______________________________________________
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



      

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

Message: 24
Date: Sat, 14 Feb 2009 08:26:01 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] freezing paraffin blocks
To: Histonet <@t> lists.utsouthwestern.edu,	Jennifer Anderson
	<janderson <@t> halozyme.com>
Message-ID: <358596.26376.qm <@t> web65713.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Not at all!
René J.

--- On Fri, 2/13/09, Jennifer Anderson <janderson <@t> halozyme.com> wrote:

From: Jennifer Anderson <janderson <@t> halozyme.com>
Subject: [Histonet] freezing paraffin blocks
To: Histonet <@t> lists.utsouthwestern.edu
Date: Friday, February 13, 2009, 5:43 PM

Hi All.

 

I've just started sectioning Bouin's fixed paraffin embedded
human/mouse/rat/pig skin again, tissues processed in a softer wax than
I'm used to, so I've been putting the blocks in the freezer at -20
prior
to sectioning.  Will such cold deleteriously affect the blocks or
tissue?

 

Also, is there any harm in cleaning the water bath with a little xylene
to remove any wax build-up?

 

Thanks a lot.

 

Jennifer M. Anderson, Scientist

Halozyme Therapeutics, Inc.

11404 Sorrento Valley Road

San Diego, CA 92121

858-704-8333

janderson <@t> halozyme.com

 

 


The information transmitted in this email is confidential and is intended only
for the person(s) or entity to which it is addressed.  Delivery of this message
to any person other than the intended recipient(s) is not intended in any way to
waive confidentiality or any applicable privilege.  Any review, retransmission,
dissemination or other use of, or taking of any action in reliance upon, this
information by individuals or entities other than the intended recipient is
prohibited by Halozyme and may be in violation of applicable laws.  If you
received this in error, please contact the sender and delete/destroy this email.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



      

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

Message: 25
Date: Sat, 14 Feb 2009 11:53:58 -0600
From: pbrunko <@t> siue.edu
Subject: [Histonet] Problem with Schiff's reagent
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <1234634038.499705367a3f6 <@t> webmail.siue.edu>
Content-Type: text/plain; charset=ISO-8859-1


Good Morning!

I'm a newcomer to the list, and I'll start by pointing out that I'm not a
histologist by any means.  I am a freshwater ecologist, and we're trying to
study mucus secretion behavior in freshwater snails.

Last Spring and last summer, we developed a process whereby we could visualize
snail mucus trails on glass slides using a periodic acid-Schiff's reagent
staining technique.  But now, in following the same protocol as used last year
for making our own Schiff's reagent, I cannot get the final solution to filter
out clear.

Recipe I'm using:
900 ml boiling water
10 grams basic fuchsin
25 ml concentrated HCl acid (12 M)
40 grams sodium metabisulfite
(this is essentially Sigma Aldrich's ratios, I think)

Let this sit for 24 to 72 hours, take 100 ml aliquot, add 0.75 - 1.0 gram
ground activated charcoal, stir for 10 minutes, filter through filter paper
then through GF/C glass fiber filter.  Last summer I got nice, clear (slightly
yellow) and very active Schiff's reagent.

But now I cannot seem to get the filtrate to be clear.  Even after 10 minutes
exposure to ground activated charcoal and filtering, the filtrate remains
bright orange to dark red and it does not seem to stain mucus trails very well.

All the reagents are the same as those used last summer (i.e., less than 7
months old; although the HCl is a bottle several years old from a different
lab).

Anyone have any troubleshooting suggestions?  I don't know the chemistry very
well, but the sodium metabisulfite is used for "decoloring" the initial
solution, right?  So is the metabisulfite not working for some reason now??

Any help/suggestions would be greatly appreciated.

Cheers -

Paul

Paul E. Brunkow, PhD
Department of Biological Sciences
Southern Illinois University Edwardsville
Edwardsville, IL    USA
-------------------------------------------------
SIUE Web Mail





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

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