[Histonet] RE: Histonet Digest, Vol 7, Issue 15

Dorothy.L.Webb <@t> HealthPartners.Com Dorothy.L.Webb <@t> HealthPartners.Com
Thu Jun 24 14:14:52 CDT 2004


The recipe for DAVIDSON's FIXATIVE is:  1500 ml. of 37& formaldehyde
                                        2250 ml. of Absolute Alcohol
                                         75 ml. of glacial acetic acid
                                        2250 ml. tap water
                                        eosin to color
We use this daily and find it works as well as all of the Penn-fix,
Disect-Aid, etc.  It fixes fatty tissue well and "pulls" the lymph nodes out
of the tissue for easier identification.  We cassette all breast biopsies
and core biopsies and put them in a rack in Davidson's, as well as fixing
the large colons and breasts in Davidsons before cassetting. 

-----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: Friday, June 11, 2004 12:04 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 7, Issue 15


Send Histonet mailing list submissions to
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Today's Topics:

   1. CPV Correction on protocol (Connie McManus)
   2. I'm sorry I can not reply immediately because I am out of the
      office, returning on Tuesday June 15, (John Ryan)
   3. RE: LKB Historange (Jurcisek, Joseph)
   4. Embedding previously frozen brain (Jo Dee Fish)
   5. Re: Embedding previously frozen brain (Gayle Callis)
   6. Perfix protocol (Boone, Linda  (HSC))
   7. Perfix  (Boone, Linda  (HSC))
   8. RE: better perfusion and inclusion---thanks to y'all
      (Charles  Scouten)
   9. best Microtomes (Tom Wells)
  10. Re: best Microtomes (EliMarGo <@t> aol.com)
  11. RE: Embedding previously frozen brain (S Ladd)
  12. long term storage of frozen sections (Instrumedics)
  13. RE: Embedding previously frozen brain (GUTIERREZ, JUAN)
  14. Histonet: Caspase-3 In Animal Tissues (Sharon E Willman)
  15. RE: Histonet: Caspase-3 In Animal Tissues (Elizabeth Chlipala)
  16. RE: Embedding previously frozen brain (Antonia Abeyta)
  17. RE: Embedding previously frozen brain (Bill Blank)
  18. (no subject) (Rush, Joyce)
  19. RE: (no subject) (Marshall Terry Dr,	Consultant Histopathologist)
  20. cd138 vendor (Martha Ward)
  21. Re: (no subject) (Vinnie Della Speranza)
  22. RE: cd138 vendor (Poteete, Jacquie A.)
  23. Re: cd138 vendor (rfail)


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

Message: 1
Date: Thu, 10 Jun 2004 11:01:15 -0600
From: "Connie McManus" <convmcm <@t> cc.usu.edu>
Subject: [Histonet] CPV Correction on protocol
To: "HISTONET" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001401c44f0c$8a540320$4a737b81 <@t> Cygnus>
Content-Type: text/plain;	charset="US-ASCII"

 
OOOPS.  I made a big boo-boo.  I stated earlier that in my CPV protocol
there was no need for Antigen retrieval.  That was only  according to
the US Biological if using their ab.  The VMRD ab reference cited calls
for Proteinase K, 5 minutes at RT.  
 
 
Connie McManus
Utah Veterinary Diagnostics Laboratory
Utah State University
Logan, UT
Phone:  435/797-1891
fax: 435/797-2805
 


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

Message: 2
Date: Thu, 10 Jun 2004 12:05:01 -0500
From: "John Ryan" <jryan <@t> sleh.com>
Subject: [Histonet] I'm sorry I can not reply immediately because I am
	out of the office, returning on Tuesday June 15,
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <s0c84e73.050 <@t> ms1.sleh.com>
Content-Type: text/plain; charset=us-ascii

I'm sorry I can not reply immediately because I am out of the office,
returning on Tuesday June 15, 2004.


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

Message: 3
Date: Thu, 10 Jun 2004 13:21:14 -0400
From: "Jurcisek, Joseph" <JurciseJ <@t> pediatrics.ohio-state.edu>
Subject: [Histonet] RE: LKB Historange
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<1366FE962DB3D311B8DA00A0C9EA048404B62A75 <@t> resms01.columbuschildrens.net>
	
Content-Type: text/plain;	charset="iso-8859-1"

I purchased my unit from a company called International Medical Equipment.
Their address information is below, I hope this helps.


	International Medical Equipment
	170 Vallecitos De Oro, Suite A
	San Marcos, CA 92069
	1-800-543-8496


Joe

Joseph A. Jurcisek
Research Associate II
Columbus Children's Research Institute
700 Children's Drive
Columbus, Ohio 43205-2696
(614) 722-2517
fax (614) 722-2007
jurcisej <@t> pediatrics.ohio-state.edu









 



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

Message: 4
Date: Thu, 10 Jun 2004 11:21:51 -0700
From: Jo Dee Fish <jfish <@t> gladstone.ucsf.edu>
Subject: [Histonet] Embedding previously frozen brain
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <v04210101bcee563d6b43@[10.40.2.112]>
Content-Type: text/plain; charset="us-ascii" ; format="flowed"

Hello Netters,
I have a question that came from an investigator at my institute:

Can mouse brain that has been snap frozen in liquid nitrogen (stored 
at -80 C degrees) be thawed and embedded in OCT for cryosectioning 
without causing excessive damage?  How can this be done?

I appreciate all of your answers and thank you all in advance.

Take care,
Jo Dee


********************************************************************** 
**********

Jo Dee Fish
Research Technologist III
Gladstone Institute of Cardiovascular Disease

Telephone: (415) 695-3720
Fax: (415) 285-5632
E-mail: jfish <@t> gladstone.ucsf.edu

Mailing address:
Gladstone Institutes
P.O. Box 419100
San Francisco, CA 94141-9100



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

Message: 5
Date: Thu, 10 Jun 2004 13:09:06 -0600
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: Re: [Histonet] Embedding previously frozen brain
To: Jo Dee Fish <jfish <@t> gladstone.ucsf.edu>,
	Histonet <@t> lists.utsouthwestern.edu
Message-ID: <3.0.6.32.20040610130906.00c09d58 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"

I don't think it will be entirely free of artifact but we have done
successful thawing/fixation then processing as described. 

To thaw, just drop the frozen brain into formalin, it will thaw while
fixing, change NBF at least once to get rid of OCT and continue to let
brain fix, then process as usual.  

At 11:21 AM 6/10/2004 -0700, you wrote:
>Hello Netters,
>I have a question that came from an investigator at my institute:
>
>Can mouse brain that has been snap frozen in liquid nitrogen (stored 
>at -80 C degrees) be thawed and embedded in OCT for cryosectioning 
>without causing excessive damage?  How can this be done?
>
>I appreciate all of your answers and thank you all in advance.
>
>Take care,
>Jo Dee
>
>
>********************************************************************** 
>**********
>
>Jo Dee Fish
>Research Technologist III
>Gladstone Institute of Cardiovascular Disease
>
>Telephone: (415) 695-3720
>Fax: (415) 285-5632
>E-mail: jfish <@t> gladstone.ucsf.edu
>
>Mailing address:
>Gladstone Institutes
>P.O. Box 419100
>San Francisco, CA 94141-9100
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 6
Date: Thu, 10 Jun 2004 14:39:29 -0500
From: "Boone, Linda  (HSC)" <Linda-Boone <@t> ouhsc.edu>
Subject: [Histonet] Perfix protocol
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<F4D83F1A4443D6459DA7E983AB5B71970D2CAD73 <@t> PARADOX.hsc.net.ou.edu>
Content-Type: text/plain;	charset="US-ASCII"

Hello All,

Does anyone happen to have the procedure to make/mix up the fixative
Perfix?  And I am also looking for the protocol for Davidson's fixative
Thanks in advance, 

Linda Boone

DMEI

Oklahoma City, OK 73104

Fax no: 405-271-3721



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

Message: 7
Date: Thu, 10 Jun 2004 14:39:36 -0500
From: "Boone, Linda  (HSC)" <Linda-Boone <@t> ouhsc.edu>
Subject: [Histonet] Perfix 
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<F4D83F1A4443D6459DA7E983AB5B71970D2CAD75 <@t> PARADOX.hsc.net.ou.edu>
Content-Type: text/plain;	charset="US-ASCII"

Hello,

Does anyone have the procedure to mix the fixative perfix? Thank you.



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

Message: 8
Date: Thu, 10 Jun 2004 14:49:37 -0500
From: "Charles  Scouten" <cwscouten <@t> myneurolab.com>
Subject: RE: [Histonet] better perfusion and inclusion---thanks to
	y'all
To: <Nancy.Walker <@t> sanofi-synthelabo.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<AF6F2F09D9DBBB4A9964C6BE5C8EC0FC539CB0 <@t> tpiserver03.Coretech-holdings.com>
	
Content-Type: text/plain;	charset="iso-8859-1"

It was my error in specifying 5% sucrose, but if you had done the Perfusion
One procedure with 10% sucrose prewash (isotonic, or slightly hypertonic),
and 300 mm Hg, you would have seen no shrinkage, and negligible ventricular
distortion. I have done it.

Please do not hesitate to try again.  You have used enough pressure to fully
wash out the red blood cells, so you have seen the clean tissue and thorough
perfusion.  You can also get the unshrunk and undistorted effects.  The
results you did get are fully predictable given the hypotonic prewash.  


Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten <@t> myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Nancy.Walker <@t> sanofi-synthelabo.com
Sent: Thursday, June 10, 2004 10:52 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] better perfusion and inclusion---thanks to y'all





More precisions:
The catheter diameter varies with animal size. For a 35 g mouse a n° 4
cathater works well (inside diameter = 0.76 mm outside = 1.22 mm).

We used 5% sucrose (saccharose was a little french slipping in) in water ,
as you had suggested. Unfortunately I didn't check out if this was in fact
isotonic,  so that is a mistake! We perfused 1 minute at 300mm Hg. The
histology looked bad, that is,  the ventricles were dilated.

Another source advised us to use Mannitol at 25% for 20 " at 110 mm Hg.
This too created ventricle dilation.

For each trial 3 mice brains were processed, coronally sliced at the
hippocampal level and stained with H/E.

yours truely,
Nancy




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

Message: 9
Date: Thu, 10 Jun 2004 16:40:04 -0700
From: "Tom Wells" <Tom_Wells <@t> bcit.ca>
Subject: [Histonet] best Microtomes
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<OFF2BBCB21.14960F9A-ON88256EAF.0080E309-88256EAF.00820345 <@t> bcit.ca>
Content-Type: text/plain; charset=us-ascii

I am the Histology Instructor at the local Institute of Technology. I am
currently in the market for manual microtomes. I have always used Leica
microtomes in the hospital and have been very happy with them. I am not
interested in motorized microtomes since repetative strain injury is not
usually an issue at this stage. What I would like is a solid,
bullet-proof, rotary microtome. These should hold up to years of student
use (abuse). I am replacing some 40 year old AO 820's that are still
working, but, only just barely. Any comments would be appreciated.
Thanks.

Tom Wells BSc, ART
Instructor
Histology/ Medical Laboratory Sciences
School of Heath Sciences
BCIT
Burnaby, BC, Canada





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

Message: 10
Date: Thu, 10 Jun 2004 23:57:18 EDT
From: EliMarGo <@t> aol.com
Subject: Re: [Histonet] best Microtomes
To: Tom_Wells <@t> bcit.ca, histonet <@t> lists.utsouthwestern.edu
Message-ID: <1dd.23b9212f.2dfa879e <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"

Hi Tom,

My facility is currently purchasing a microtome for our lab. The one that we

decided on in the end is the Microm HM 310. We are a small pathology lab in 
Los Angeles, CA and have no need for motorized microtomes either. 

The other microtome we were considering was the Leica 2125 RT and both
models 
are very much similar. In fact, the few differences stem from the different 
providers not the machine itself. One is from McBain Instruments and the
other 
is provided by Mikkron. 
If you would like, I can fax you the quotes that were given to me by each 
company. On them you will find brief descriptions of each model. Let me know
if 
you are interested.


Sincerely,

Elisa Gonzalez
Research Tech II
Doheny Eye Institute
Pathology Department


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

Message: 11
Date: Fri, 11 Jun 2004 07:58:49 -0400
From: "S Ladd" <sladd <@t> hsc.usf.edu>
Subject: RE: [Histonet] Embedding previously frozen brain
To: "Jo Dee Fish" <jfish <@t> gladstone.ucsf.edu>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <EDEILFIMBEHLDJPKFIPIEEINCBAA.sladd <@t> hsc.usf.edu>
Content-Type: text/plain;	charset="us-ascii"

Why do you want to thaw it if you are putting it in OCT and cryosectioning??
Sharron

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Jo Dee
Fish
Sent: Thursday, June 10, 2004 2:22 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Embedding previously frozen brain


Hello Netters,
I have a question that came from an investigator at my institute:

Can mouse brain that has been snap frozen in liquid nitrogen (stored 
at -80 C degrees) be thawed and embedded in OCT for cryosectioning 
without causing excessive damage?  How can this be done?

I appreciate all of your answers and thank you all in advance.

Take care,
Jo Dee


********************************************************************** 
**********

Jo Dee Fish
Research Technologist III
Gladstone Institute of Cardiovascular Disease

Telephone: (415) 695-3720
Fax: (415) 285-5632
E-mail: jfish <@t> gladstone.ucsf.edu

Mailing address:
Gladstone Institutes
P.O. Box 419100
San Francisco, CA 94141-9100

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



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

Message: 12
Date: Fri, 11 Jun 2004 09:42:39 -0400
From: "Instrumedics" <info <@t> instrumedics.com>
Subject: [Histonet] long term storage of frozen sections
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <014201c44fba$13a29530$6401a8c0 <@t> INSTRUMEDICS22>
Content-Type: text/plain;	charset="iso-8859-1"

We use the Instrumedics Protective Oil to coat the block which prevents
dehydration for at least a year in an -80C freezer.
Bernice
----- Original Message ----- 
From: <histonet-request <@t> lists.utsouthwestern.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, June 10, 2004 1:00 PM
Subject: Histonet Digest, Vol 7, Issue 14


> Send Histonet mailing list submissions to
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>
> 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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> than "Re: Contents of Histonet digest..."
>


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


> Today's Topics:
>
>    1. Cryostorage -> how to prevent desiccation ? (knibbelmann <@t> web.de)
>    2. Re: Cryostorage -> how to prevent desiccation ? (Helen Fedor)
>    3. RE:  LKB Historange 2218 (Lett, Jaclynn)
>    4. Histology/Electron Microscopy position (Breisch, Eric)
>    5. Re: charge for cryostat use (Ford Royer)
>    6. RE: Toe nails !!!!! (Thom Jensen)
>    7. S100A6 (Patti Loykasek)
>    8. Re: Reconditioned Leica Autostainer or Sikora Bach Stainer
>       (Don.Birgerson <@t> leica-microsystems.com)
>    9. Centralized Accessioning (Bonnie J. McMahill)
>   10. Kerrie Thomson/MELB/AU/BAYER is out of the office.
>       (kerrie.thomson.kt <@t> bayer-ag.de)
>   11. anti-CDV, Neospora and TGE (path_kimron)
>   12. caspase -12 (Inga Hansson)
>   13. better perfusion and inclusion---thanks to y'all
>       (Nancy.Walker <@t> sanofi-synthelabo.com)
>   14. IHC protocols for Parvovirus Cat and dog (Lab BioVet AB)
>   15. Re: IHC protocols for Parvovirus Cat and dog (Greg Dobbin)
>   16. Re: IHC protocols for Parvovirus Cat and dog (Jan Shivers)
>   17. Re: anti-CDV, Neospora and TGE (Jan Shivers)
>   18. RE: better perfusion and inclusion---thanks to y'all
>       (Charles  Scouten)
>   19. RE: better perfusion and inclusion---thanks to y'all
>       (Nancy.Walker <@t> sanofi-synthelabo.com)
>   20. Sudan Black B for lipofuscin? (McCollough, Carol)
>   21. telepathology??? (Lesley S. Bechtold)
>


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


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





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

Message: 13
Date: Fri, 11 Jun 2004 08:47:45 -0500
From: "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org>
Subject: RE: [Histonet] Embedding previously frozen brain
To: "Jo Dee Fish" <jfish <@t> gladstone.ucsf.edu>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<A61F23E937E4DA488E0C0F60093843D901E49735 <@t> ccetxm030.echristus.net>
Content-Type: text/plain;	charset="iso-8859-1"

The way we thawed specimens at a lab I used to work for, is by putting the
frozen tissue in refrigerated PBS for a few minutes.  We then patted it dry
on a piece of filter paper and proceeded with the OCT embedding.  Of course
that's if you want your tissue completely surrounded by OCT.  If not, you
can just mount it directly in OCT from the -80.

Juan C. Gutierrez, HT(ASCP)
Histology Laboratory Supervisor
Christus Santa Rosa Hospital
333 N. Santa Rosa Ave.
San Antonio, TX  78207
(210)704-2533


-----Original Message-----
From: Jo Dee Fish [mailto:jfish <@t> gladstone.ucsf.edu]
Sent: Thursday, June 10, 2004 1:22 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Embedding previously frozen brain


Hello Netters,
I have a question that came from an investigator at my institute:

Can mouse brain that has been snap frozen in liquid nitrogen (stored 
at -80 C degrees) be thawed and embedded in OCT for cryosectioning 
without causing excessive damage?  How can this be done?

I appreciate all of your answers and thank you all in advance.

Take care,
Jo Dee


********************************************************************** 
**********

Jo Dee Fish
Research Technologist III
Gladstone Institute of Cardiovascular Disease

Telephone: (415) 695-3720
Fax: (415) 285-5632
E-mail: jfish <@t> gladstone.ucsf.edu

Mailing address:
Gladstone Institutes
P.O. Box 419100
San Francisco, CA 94141-9100

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



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

Message: 14
Date: Fri, 11 Jun 2004 08:47:49 -0500
From: Sharon E Willman <sharon.willman <@t> bms.com>
Subject: [Histonet] Histonet: Caspase-3 In Animal Tissues
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <40C9B805.4872C8B <@t> bms.com>
Content-Type: text/plain; charset=us-ascii

Hi,
Has anyone has had success doing  R&D caspase-3 IHC in animal
tissues?  Any information you might be able to share would be
most appreciated.

Thanks in advance.

Sharon Willman




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

Message: 15
Date: Fri, 11 Jun 2004 08:10:46 -0600
From: "Elizabeth Chlipala" <lizchlipala <@t> premierhistology.com>
Subject: RE: [Histonet] Histonet: Caspase-3 In Animal Tissues
To: "'Sharon E Willman'" <sharon.willman <@t> bms.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000401c44fbd$e74c7f60$74d48a80 <@t> LIZ>
Content-Type: text/plain;	charset="us-ascii"

Sharon

I have not used R&D's caspase-3, but I have worked with cell signaling
technologies caspase-3 in human, rat and mouse specimens.  This antibody
works well with high pH retrieval.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)
Premier Histology Laboratory, LLC
P.O. Box 18592 
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
lizchlipala <@t> premierhistology.com
www.premierhistology.com
 
Ship to Address:
Premier Histology Laboratory
University of Colorado
MCBD, Room A3B40
Boulder, Colorado 80309
 
_________________________________ 
This email and any files transmitted with it may contain PRIVILEGED or
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sharon E
Willman
Sent: Friday, June 11, 2004 6:48 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Histonet: Caspase-3 In Animal Tissues

Hi,
Has anyone has had success doing  R&D caspase-3 IHC in animal
tissues?  Any information you might be able to share would be
most appreciated.

Thanks in advance.

Sharon Willman


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




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

Message: 16
Date: Fri, 11 Jun 2004 08:15:45 -0600
From: "Antonia Abeyta" <ALAbeyta <@t> salud.unm.edu>
Subject: RE: [Histonet] Embedding previously frozen brain
To: <juan.gutierrez <@t> christushealth.org>,<jfish <@t> gladstone.ucsf.edu>,
	<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s0c96a48.093 <@t> salud.unm.edu>
Content-Type: text/plain; charset=US-ASCII

I personally find it easier to mount it directly on OCT from the -80. 
In my experience, if you embed the tissue, the sections are more likely
to curl.

Antonia


>>> "GUTIERREZ, JUAN" <juan.gutierrez <@t> christushealth.org> 6/11/2004
7:47:45 AM >>>
The way we thawed specimens at a lab I used to work for, is by putting
the frozen tissue in refrigerated PBS for a few minutes.  We then patted
it dry on a piece of filter paper and proceeded with the OCT embedding. 
Of course that's if you want your tissue completely surrounded by OCT. 
If not, you can just mount it directly in OCT from the -80.

Juan C. Gutierrez, HT(ASCP)
Histology Laboratory Supervisor
Christus Santa Rosa Hospital
333 N. Santa Rosa Ave.
San Antonio, TX  78207
(210)704-2533


-----Original Message-----
From: Jo Dee Fish [mailto:jfish <@t> gladstone.ucsf.edu] 
Sent: Thursday, June 10, 2004 1:22 PM
To: Histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] Embedding previously frozen brain


Hello Netters,
I have a question that came from an investigator at my institute:

Can mouse brain that has been snap frozen in liquid nitrogen (stored 
at -80 C degrees) be thawed and embedded in OCT for cryosectioning 
without causing excessive damage?  How can this be done?

I appreciate all of your answers and thank you all in advance.

Take care,
Jo Dee


**********************************************************************

**********

Jo Dee Fish
Research Technologist III
Gladstone Institute of Cardiovascular Disease

Telephone: (415) 695-3720
Fax: (415) 285-5632
E-mail: jfish <@t> gladstone.ucsf.edu 

Mailing address:
Gladstone Institutes
P.O. Box 419100
San Francisco, CA 94141-9100

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

Message: 17
Date: Fri, 11 Jun 2004 09:16:28 -0500
From: Bill Blank <bill501 <@t> mindspring.com>
Subject: RE: [Histonet] Embedding previously frozen brain
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <p06010207bcef6e1d2c9f@[63.153.12.39]>
Content-Type: text/plain; charset="us-ascii" ; format="flowed"

At 7:58 AM -0400 6/11/04, S Ladd wrote:
>Why do you want to thaw it if you are putting it in OCT and
cryosectioning??
>Sharron

My thoughts exactly. Just place in the cryostat and let it warm up to 
the cryostat temp and slice away. We always did brain frozen sections 
this way.
-- 
______________
Bill Blank, MD
Heartland Lab, Inc



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

Message: 18
Date: Fri, 11 Jun 2004 09:37:17 -0500
From: "Rush, Joyce" <Joyce.Rush <@t> sjmcmn.org>
Subject: [Histonet] (no subject)
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<494BA9CA5AA74C48B644FCA2B8BECEC844D75B <@t> sjw3smail.SJMCMN.ORG>
Content-Type: text/plain;	charset="iso-8859-1"

I am a Lab Manager, not a Histotech, and I need your help. We are a general
pathology practice and need to know what kind of processing cycles others
use for small bx's.  Currently our lab processes all tissues together, large
and small, and therefore has much trouble with over processed bx's.  
 
I would very much appreciate your guidance so that I can help our histology
area move forward.
 
Thanks so much!
 
Joyce

Joyce A. Rush, BS, MT(ASCP) 
Laboratory Manager 
St Joseph's Medical Center 
523 North Third Street 
Brainerd, MN  56401 
Office:218-828-7500   Fax:  218-828-7510 

 




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thereof and destroy/delete the message. Please note that any views or
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check this email and any attachments for the presence of viruses. The
Medical Center accepts no liability for any damage caused by any virus
transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street,
Brainerd, MN 56401

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

Message: 19
Date: Fri, 11 Jun 2004 15:48:02 +0100
From: "Marshall Terry Dr,	Consultant Histopathologist"
	<Terry.Marshall <@t> rothgen.nhs.uk>
Subject: RE: [Histonet] (no subject)
To: "Rush, Joyce" <Joyce.Rush <@t> sjmcmn.org>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<FE2DB935F8BBB546B8A1BBF3459C5A1F2CC6E7 <@t> LIL.xRothGen.nhs.uk>
Content-Type: text/plain;	charset="iso-8859-1"

The concept of over-processed tissue crops up regularly.
Is there general agreement:

a/	That over-processed biopsies exist
b/	That they show some characteristic(s) by which they can be regularly
recognised
c/	That the cycle or some part thereof needs to be reduced in time to
correct the features seen in an affirmative answer 	to b/?

Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
 Consultant Pathologist
 Rotherham General Hospital
 South Yorkshire
 England
        terry.marshall <@t> rothgen.nhs.uk

-----Original Message-----
From: Rush, Joyce [mailto:Joyce.Rush <@t> sjmcmn.org]
Sent: 11 June 2004 15:37
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] (no subject)


I am a Lab Manager, not a Histotech, and I need your help. We are a general
pathology practice and need to know what kind of processing cycles others
use for small bx's.  Currently our lab processes all tissues together, large
and small, and therefore has much trouble with over processed bx's.  
 
I would very much appreciate your guidance so that I can help our histology
area move forward.
 
Thanks so much!
 
Joyce

Joyce A. Rush, BS, MT(ASCP) 
Laboratory Manager 
St Joseph's Medical Center 
523 North Third Street 
Brainerd, MN  56401 
Office:218-828-7500   Fax:  218-828-7510 

 




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necessarily represent those of the company. Finally, the recipient should
check this email and any attachments for the presence of viruses. The
Medical Center accepts no liability for any damage caused by any virus
transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street,
Brainerd, MN 56401
_______________________________________________
Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 20
Date: Fri, 11 Jun 2004 12:29:08 -0400
From: "Martha Ward" <mward <@t> wfubmc.edu>
Subject: [Histonet] cd138 vendor
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<61135F0455D33347B5AAE209B903A304076A4E9B <@t> EXCHVS2.medctr.ad.wfubmc.edu>
	
Content-Type: text/plain;	charset="us-ascii"

Can anyone recommend a vendor, etc. for CD138 (syndecan-1) that works
well on either the Ventana NexEs or the Dako stainer?  Thanks in
advance!
 
Martha Ward
Wake Forest University Baptist Medical Center


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

Message: 21
Date: Fri, 11 Jun 2004 12:30:21 -0400
From: "Vinnie Della Speranza" <dellav <@t> musc.edu>
Subject: Re: [Histonet] (no subject)
To: <histonet <@t> lists.utsouthwestern.edu>, <Joyce.Rush <@t> sjmcmn.org>
Message-ID: <s0c9a5eb.026 <@t> cl.musc.edu>
Content-Type: text/plain; charset=US-ASCII

Excellent question. You are quite correct that processing all tissues
together will create avoidable problems.

generally, needle biopsy or small endoscopic specimens can be processed with
as little as ten-fifteen minutes in each alcohol and xylene step. if your
processor has three or more paraffin steps, you can use ten or fifteen
minutes in each. Keep in mind that the first paraffin is frequently
contaminated with xylene so subsequent exposure of the tissues to "clean"
paraffin is key to good infiltration of the tissuesl We turn our "rush"
biopsies around in about an hour on the processor with about 5 minutes in
each step and still get very good results. It is important of course that
you have a schedule in place for the regular freshening or changing of
processing solutions.

Let me make you aware of a relatively new (year 2) proficiency program that
you may want to consider for your laboratory. Its called Histo-QIP offered
jointly by the CAP and the NSH. It assesses the quality of your histologic
preparations and stains using a variety of exercises the subscribing labs
submit for evaluation. You receive an assessment and detailed educational
material back that can be used to educate staff. Participation gives you a
good idea of what aspects of your operation may need corrective action AND
steps on how to achieve the quality you want. I participate on the
Histotechnology committee that developed the Histo-QIP program for the CAP
but I also enroll my lab because of my strong feelings for the program

biopsies processed on long cycles normally reserved for larger excisions
will be at risk for having protein-bound water removed, the effects of which
can be quite dramatic. These typically include pyknotic nuclei with very
poor chromatin detail. they are very dark and appear to be overstained..
overprocessed tissues may also be very difficult to cut.

Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue  Suite 309
Charleston, SC 29425
Ph: 843-792-6353
fax: 843-792-8974

>>> "Rush, Joyce" <Joyce.Rush <@t> sjmcmn.org> 06/11/04 10:37AM >>>
I am a Lab Manager, not a Histotech, and I need your help. We are a general
pathology practice and need to know what kind of processing cycles others
use for small bx's.  Currently our lab processes all tissues together, large
and small, and therefore has much trouble with over processed bx's.  
 
I would very much appreciate your guidance so that I can help our histology
area move forward.
 
Thanks so much!
 
Joyce

Joyce A. Rush, BS, MT(ASCP) 
Laboratory Manager 
St Joseph's Medical Center 
523 North Third Street 
Brainerd, MN  56401 
Office:218-828-7500   Fax:  218-828-7510 

 




DISCLAIMER: This email and any files transmitted with it are confidential
and intended solely for the use of the individual or entity to whom they are
addressed. If you have received this email in error please notify the sender
thereof and destroy/delete the message. Please note that any views or
opinions presented in this email are solely those of the Author and do not
necessarily represent those of the company. Finally, the recipient should
check this email and any attachments for the presence of viruses. The
Medical Center accepts no liability for any damage caused by any virus
transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street,
Brainerd, MN 56401
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




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

Message: 22
Date: Fri, 11 Jun 2004 11:44:01 -0500
From: "Poteete, Jacquie A." <japoteete <@t> saintfrancis.com>
Subject: RE: [Histonet] cd138 vendor
To: 'Martha Ward' <mward <@t> wfubmc.edu>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<D423BBB85E91D411A38D42003000996506973E8C <@t> mailnt1.saintfrancis.com>
Content-Type: text/plain

We use DAKO's CD138, catalog number M7228, and are very happy with it.

Jacquie Poteete MT(ASCP)QIHC
Lead Technologist, IHC Laboratory
Saint Francis Hospital, Tulsa, OK
japoteete <@t> saintfrancis.com

> -----Original Message-----
> From:	Martha Ward [SMTP:mward <@t> wfubmc.edu]
> Sent:	Friday, June 11, 2004 11:29 AM
> To:	histonet <@t> lists.utsouthwestern.edu
> Subject:	[Histonet] cd138 vendor
> 
> Can anyone recommend a vendor, etc. for CD138 (syndecan-1) that works
> well on either the Ventana NexEs or the Dako stainer?  Thanks in
> advance!
>  
> Martha Ward
> Wake Forest University Baptist Medical Center
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> 
> 
********* Email Confidentiality Statement ********* 
Visit http://www.saintfrancis.com/emailconf.asp



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

Message: 23
Date: Fri, 11 Jun 2004 12:48:42 -0400
From: rfail <rfail <@t> toolkitmail.com>
Subject: Re: [Histonet] cd138 vendor
To: Martha Ward <mward <@t> wfubmc.edu>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <40c9e26a.f8.7092.1911454150 <@t> toolkitmail.com>

DAKO

 Can anyone recommend a vendor, etc. for CD138 (syndecan-1)
> that works well on either the Ventana NexEs or the Dako
> stainer?  Thanks in advance!
>
> Martha Ward
> Wake Forest University Baptist Medical Center
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

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End of Histonet Digest, Vol 7, Issue 15
***************************************

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