[Histonet] RE: Histonet Digest, Vol 35, Issue 28, heat antigen retrieval methods

From:"Goodwin, Diana"

Peter:

Check out the June 2006 issue of the Medical Laboratory Observer(MLO)-www=2Emlo-online.com.  The cover story was on AR.

Diana Goodwin
Pennsylvania Hospital
Philadelphia, PA 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu
Sent: Tuesday, October 17, 2006 1:09 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 35, Issue 28

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Today's Topics:

   1. Milestone Pathos vs Sakura Xpress (Gregor Arlt)
   2. RE: Milestone Pathos vs Sakura Xpress (Jes Strong)
   3. Re: Milestone Pathos vs Sakura Xpress (Rene J Buesa)
   4. RE: Milestone Pathos vs Sakura Xpress
      (Bartlett, Jeanine (CDC/CCID/NCZVED))
   5. seeking hiotology position in DC area (Steven Wilkes)
   6. Re: Vison BioSystems - Peloris (Anthony Reilly)
   7. Hello,	fellow histotechies!!!  This is my first time posing a
      question - so be gentle.  Our patholog (Yvonne Jones)
   8. RE: Milestone Pathos vs Sakura Xpress
      (Bartlett, Jeanine (CDC/CCID/NCZVED))
   9. RE: Hello, fellow histotechies!!! This is my first	time
      posing a question - so be gentle. Our patholog
      (rdavis4@rdg.boehringer-ingelheim.com)
  10. RE: Hello,	fellow histotechies!!! This is my first time
      posing a question - so	be gentle. Our patholog (soofia siddiqui)
  11. heat antigen retrieval methods (Peter Rippstein)
  12. problems with mouse brain fixation (Martina Urbanek)
  13. Phospho-S6 Ribosomal Protein (Goodwin, Diana)


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

Message: 1
Date: Mon, 16 Oct 2006 14:53:33 -0500
From: "Gregor Arlt" 
Subject: [Histonet] Milestone Pathos vs Sakura Xpress
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; format=flowed

Dear Histonetters,

I have heard the Pathos would use 600 Watts for the microwave processing. I can't imagine that this is right. In my opinion this would destroy the RNS structure of any tissue. On the other hand I heard tissues processed in the Xpress would be easy to use for molecularbiological investigations.

Has anybody experience with those instruments, or know anybody the power of the microwave of the instruments.

Thanks for the help Frank

_________________________________________________________________
Get FREE company branded e-mail accounts and business Web site from Microsoft Office Live http://clk.atdmt.com/MRT/go/mcrssaub0050001411mrt/direct/01/




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

Message: 2
Date: Mon, 16 Oct 2006 15:43:55 -0500
From: "Jes Strong" 
Subject: RE: [Histonet] Milestone Pathos vs Sakura Xpress
To: 
Message-ID: <00a501c6f163$cc5b0010$0200a8c0@Jes>
Content-Type: text/plain;	charset="us-ascii"

Dear Mr. Arlt,

If you would like to contact Milestone directly, we would be happy to explain the principles of Milestone's microwave processors including magnetron output and how it is dynamically regulated by software to adjust to each specific load. These are not questions that users would, or should be expected to be able to answer for you satisfactorily.  

Jes Strong
Western Region Sales Manager
Milestone Medical
(203) 925-4240 (Office)
(847) 323-8373 (Cell)
(847) 655-6009 (Fax)
jes@milestonemed.com
 
www.milestonemed.com
 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gregor Arlt
Sent: Monday, October 16, 2006 2:54 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Milestone Pathos vs Sakura Xpress

Dear Histonetters,

I have heard the Pathos would use 600 Watts for the microwave processing. I can't imagine that this is right. In my opinion this would destroy the RNS structure of any tissue. On the other hand I heard tissues processed in the Xpress would be easy to use for molecularbiological investigations.

Has anybody experience with those instruments, or know anybody the power of the microwave of the instruments.

Thanks for the help Frank

_________________________________________________________________
Get FREE company branded e-mail accounts and business Web site from Microsoft Office Live http://clk.atdmt.com/MRT/go/mcrssaub0050001411mrt/direct/01/


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

Message: 3
Date: Mon, 16 Oct 2006 14:19:18 -0700 (PDT)
From: Rene J Buesa 
Subject: Re: [Histonet] Milestone Pathos vs Sakura Xpress
To: Gregor Arlt ,
	histonet@lists.utsouthwestern.edu
Message-ID: <20061016211918.92177.qmail@web61214.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Frank:
  The temperature produced by the magnetron is "controlled" by the temperature probe  adjusted to the processing protocol; you can have 1200W and a working temperature of 50C so wattage in itself is not a deletereous agent just provides the capability of getting to high temperatures quickly, if needed..
  Not all technologies are alike and the Xpress uses 60W continuously in the first 2 chambers. PATHOS is pure microwave technology and Xpress is a blend of MW and conventional technology.
  In the Xpress the first 2 chambers are identical and  using MW technology and bubble agitation. The other 2 are just 2 conventional retorts with convection heat and vacuum capabilities, operated at 65C.
  The molecular integrity does not relate to the process but to the tissue fixation. Formalin greatly prevents RNA studies but any alcoholic fixative like Kryofix, BoonFix or the propietary by Sakura (UMFix) will preserve the macromolecules either if the tissue is going to be processed with MW or conventional technology. Any good alcoholic fixative containin PEG also will preserve the macromolecules.
  Both PATHOS and Xpress are "walk away" instruments but Xpress allows for the continuous addition of up to 30 cassettes every 15 minutes, for an overall work flow of 120 cassettes after 105 minutes, and 30 more every 15 minutes afterwards. The limit is the thickness of the sections (have to be 1.5mm thick) and some tissues have to be previously fixed from 4 to 4=2E5 hours before processing.
  PATHOS can process tissues of up to 5mm at a rate of 210 cassettes/4 hours (fixation included) for thick tissue slices or 210 cassettes/1 hour for small biopsies.
  Hope this information will help you!
  Ren J.

Gregor Arlt  wrote:
  Dear Histonetters,

I have heard the Pathos would use 600 Watts for the microwave processing. I can't imagine that this is right. In my opinion this would destroy the RNS structure of any tissue. On the other hand I heard tissues processed in the Xpress would be easy to use for molecularbiological investigations.

Has anybody experience with those instruments, or know anybody the power of the microwave of the instruments.

Thanks for the help Frank

_________________________________________________________________
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Message: 4
Date: Mon, 16 Oct 2006 18:46:57 -0400
From: "Bartlett, Jeanine \(CDC/CCID/NCZVED\)" 
Subject: RE: [Histonet] Milestone Pathos vs Sakura Xpress
To: "Gregor Arlt" ,
	
Message-ID:
	
Content-Type: text/plain;	charset="UTF-8"

Frank,
 
I have no experience with the Pathos but our lab does have the Sakura Xpress.  I am attaching a PDF of the brochure for the equipment.  Page 4 explains how the microwave itself operates (60 watts).  One thing I want to add to the previous response is that the 1.5 mm thickness for the Xpress is not mandatory.  There is allowance for thicker specimens but the processing time is then lengthened. But I for one like having an excuse to make pathologists gross properly to begin with. :)
 
Jeanine Bartlett
CDC, Atlanta

	-----Original Message----- 
	From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gregor Arlt 
	Sent: Mon 10/16/2006 3:53 PM 
	To: histonet@lists.utsouthwestern.edu 
	Cc: 
	Subject: [Histonet] Milestone Pathos vs Sakura Xpress
	
	

	Dear Histonetters,
	
	I have heard the Pathos would use 600 Watts for the microwave processing. I
	can't imagine that this is right. In my opinion this would destroy the RNS
	structure of any tissue. On the other hand I heard tissues processed in the
	Xpress would be easy to use for molecularbiological investigations.
	
	Has anybody experience with those instruments, or know anybody the power of
	the microwave of the instruments.
	
	Thanks for the help Frank
	
	_________________________________________________________________
	Get FREE company branded e-mail accounts and business Web site from
	Microsoft Office Live
	http://clk.atdmt.com/MRT/go/mcrssaub0050001411mrt/direct/01/
	
	
	_______________________________________________
	Histonet mailing list
	Histonet@lists.utsouthwestern.edu
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
	


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

Message: 5
Date: Mon, 16 Oct 2006 22:15:25 -0400
From: "Steven Wilkes" 
Subject: [Histonet] seeking hiotology position in DC area
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<8e5827cf0610161915w1f450489mb6aec23f432807ae@mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Hello

I am seeking a histology position in the greater DC area.  I have a MA in biology, 2+ years of histology and immunohistochemistry experience, as well as a bit histology teaching.  Thank you

Steven


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

Message: 6
Date: Tue, 17 Oct 2006 12:37:52 +1000
From: "Anthony Reilly" 
Subject: Re: [Histonet] Vison BioSystems - Peloris
To: ,
Message-ID: 
Content-Type: text/plain; charset="us-ascii"

Hi Richard

My peloris was part of the very first relase of the instrument.  As a result there were some initial minor problems which have since been rectified.  As I said these problems were minor and never in any way affected the processing of our tissue.

It now runs very well and has had a positive impact on our laboratory.
The instrument has a very powerful mixing ability which not only improves penetration but guarantees even heating on the steps where heat is utilised giving shorter processing times even for fatty tissue. This is aided by using one of their range of specimen baskets which separates each cassette individually allowing better flow of solution to each specimen.

Our laboratory services both heart and lung transplant units requiring us to run numeroous short cycles throughout the day.  The improved processing combined with a rapid clean cycle means that the one dual retort peloris can do the work of 3-4 of our prevoius instruments.

Examples of our improved times include:

Fatty tissue 18h to 14h
Routine 12h to 9h
Small Biopsy 2h to 1 h

This has also had an impact on IHC as the small biopsies that are required urgently can be given an extra 1h in formalin and still be completed in the same time as the previous protocol.  With the faster processing some tissues such as lletz biopsies need to be processed on shorter cycles to avoid hardening of the tissue.  According to the manufacturer these times can be reduced further by substituting xylene with isopropanol but I have not tried that so cannot comment.

regards



Tony Reilly
Chief Scientist
Anatomical Pathology
QHPS-Prince Charles Hospital
Rode Rd Chermside Q 4032
Australia
Ph: 07 3139 4543
Fax: 07 3193 4546
tony_reilly@health.qld.gov.au


>>> "Richard Cartun"  10/15/06 11:46 pm >>>
Anyone out there using Vison BioSystems' "Peloris" for tissue processing?  If so, what has been your experience?  Thank you.

Richard

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


                                        


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

Message: 7
Date: Tue, 17 Oct 2006 02:43:21 -0400
From: "Yvonne Jones" 
Subject: [Histonet] Hello,	fellow histotechies!!!  This is my first
	time posing a	question - so be gentle.  Our patholog
To: 
Message-ID: <453443490200004F00000225@GWGATE1.ahm.com>
Content-Type: text/plain;	charset="US-ASCII"

Hello, fellow histotechies!!!  This is my first time posing a question -
so be gentle.  Our pathologists have recently requested that we begin
testing an antibody, p52.  I have had a bear of a time finding any
information about this antibody!!!  All I could find was one article
on-line, and I am still having a problem finding info and the antibody
itself.  

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





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

Message: 8
Date: Tue, 17 Oct 2006 05:49:06 -0400
From: "Bartlett, Jeanine \(CDC/CCID/NCZVED\)" 
Subject: RE: [Histonet] Milestone Pathos vs Sakura Xpress
To: "Bartlett, Jeanine \(CDC/CCID/NCZVED\)" ,	"Gregor
	Arlt" ,
	
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

Apparently the PDF was too large for Histonet to accept.  You can go to
the Sakura website and click on Xpress and open the brochure if you need
these details.

Jeanine 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Bartlett, Jeanine (CDC/CCID/NCZVED)
Sent: Monday, October 16, 2006 6:47 PM
To: Gregor Arlt; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Milestone Pathos vs Sakura Xpress

Frank,
 
I have no experience with the Pathos but our lab does have the Sakura
Xpress.  I am attaching a PDF of the brochure for the equipment.  Page 4
explains how the microwave itself operates (60 watts).  One thing I want
to add to the previous response is that the 1.5 mm thickness for the
Xpress is not mandatory.  There is allowance for thicker specimens but
the processing time is then lengthened. But I for one like having an
excuse to make pathologists gross properly to begin with. :)
 
Jeanine Bartlett
CDC, Atlanta

	-----Original Message----- 
	From: histonet-bounces@lists.utsouthwestern.edu on behalf of
Gregor Arlt 
	Sent: Mon 10/16/2006 3:53 PM 
	To: histonet@lists.utsouthwestern.edu 
	Cc: 
	Subject: [Histonet] Milestone Pathos vs Sakura Xpress
	
	

	Dear Histonetters,
	
	I have heard the Pathos would use 600 Watts for the microwave
processing. I
	can't imagine that this is right. In my opinion this would
destroy the RNS
	structure of any tissue. On the other hand I heard tissues
processed in the
	Xpress would be easy to use for molecularbiological
investigations.
	
	Has anybody experience with those instruments, or know anybody
the power of
	the microwave of the instruments.
	
	Thanks for the help Frank
	
	
_________________________________________________________________
	Get FREE company branded e-mail accounts and business Web site
from
	Microsoft Office Live
	http://clk.atdmt.com/MRT/go/mcrssaub0050001411mrt/direct/01/
	
	
	_______________________________________________
	Histonet mailing list
	Histonet@lists.utsouthwestern.edu
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
	






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

Message: 9
Date: Tue, 17 Oct 2006 07:46:17 -0400
From: rdavis4@rdg.boehringer-ingelheim.com
Subject: RE: [Histonet] Hello, fellow histotechies!!! This is my first
	time posing a question - so be gentle. Our patholog
To: yjones2@csmlab.com,	histonet@lists.utsouthwestern.edu
Message-ID:
	<83BA2D3D42947D48BDAA449453644ABE0865B1@RDGEXM01.am.boehringer.com>
Content-Type: text/plain; charset=us-ascii

Yvonne,

Google p52 and lots of hits come up.  Also, check out www.abcam.com.  They
have p52 as a rabbit polyclonal.

Rebecca A. Davis, A.A.S., NYS LVT, HT (ASCP) 
Toxicology, Histopathology Lab 
Boehringer-Ingelheim Pharmaceuticals, Inc. 
rdavis4@rdg.boehringer-ingelheim.com 
203-798-5448 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yvonne Jones
Sent: Tuesday, October 17, 2006 2:43 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Hello, fellow histotechies!!! This is my first time
posing a question - so be gentle. Our patholog

Hello, fellow histotechies!!!  This is my first time posing a question -
so be gentle.  Our pathologists have recently requested that we begin
testing an antibody, p52.  I have had a bear of a time finding any
information about this antibody!!!  All I could find was one article
on-line, and I am still having a problem finding info and the antibody
itself.  

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



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





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

Message: 10
Date: Tue, 17 Oct 2006 07:39:31 -0700 (PDT)
From: soofia siddiqui 
Subject: RE: [Histonet] Hello,	fellow histotechies!!! This is my first
	time posing a question - so	be gentle. Our patholog
To: rdavis4@rdg.boehringer-ingelheim.com, yjones2@csmlab.com,
	histonet@lists.utsouthwestern.edu
Message-ID: <20061017143931.16702.qmail@web39513.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Go to Google and go check for Biocompare.com. Biocompare is a very good source to search for any bio products. I  have searched and found all of the antibodies, that Dako has discontinued, through this web site. Good luck!
  Soofia

rdavis4@rdg.boehringer-ingelheim.com wrote:
  Yvonne,

Google p52 and lots of hits come up. Also, check out www.abcam.com. They
have p52 as a rabbit polyclonal.

Rebecca A. Davis, A.A.S., NYS LVT, HT (ASCP) 
Toxicology, Histopathology Lab 
Boehringer-Ingelheim Pharmaceuticals, Inc. 
rdavis4@rdg.boehringer-ingelheim.com 
203-798-5448 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yvonne Jones
Sent: Tuesday, October 17, 2006 2:43 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Hello, fellow histotechies!!! This is my first time
posing a question - so be gentle. Our patholog

Hello, fellow histotechies!!! This is my first time posing a question -
so be gentle. Our pathologists have recently requested that we begin
testing an antibody, p52. I have had a bear of a time finding any
information about this antibody!!! All I could find was one article
on-line, and I am still having a problem finding info and the antibody
itself. 

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



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



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Message: 11
Date: Tue, 17 Oct 2006 11:36:56 -0400
From: "Peter Rippstein" 
Subject: [Histonet] heat antigen retrieval methods
To: 
Message-ID: 
Content-Type: text/plain; charset="us-ascii"

Hello Histonetters,

Our lab is in need of some information in regards  heat antigen
retrieval methods. Has anyone done a comparison in terms of results &
cost effectiveness obtained from microwave irradiation vs pressure
cooking and steam heating methods. Any recommendations would be
appreciated. Many thanks.
Peter
 

Peter Rippstein  ART, MLT
Core Pathology Laboratory
Rm H2102
University of Ottawa Heart Institute
40 Ruskin Street
Ottawa, Ontario
Canada, K1Y 4W7

Tel: (613) 761-5282
Fax: (613) 761-5281
Email: prippstein@ottawaheart.ca






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

Message: 12
Date: Tue, 17 Oct 2006 17:51:41 +0200
From: Martina Urbanek 
Subject: [Histonet] problems with mouse brain fixation
To: histonet@lists.utsouthwestern.edu
Message-ID: <1161100301.4534fc0d23f8a@web-mail1.uibk.ac.at>
Content-Type: text/plain; charset=ISO-8859-1

Hello everybody on histonet,

we have some problems with handling mouse (21 and 90 days old) and rat brains.
We do mouse brain perfusion using 4% paraformaldehyde in PBS, pH 7.4. We use
gravity for perfusion for about 30 minutes (volume about 80 ml) and after
perfusion we leave the brains over night in the same fixative. Then we process
in a Shandon tissue processor (70% Alcohol, 80% Alcohol, 95% Alcohol, 3 changes
100% alcohol, 3 changes xylene, 2 changes paraffin 56C; time depends on size
of tissue). Now we have the problem that some brains are too hard and some also
seem to shrink more than others, when we cut them and put them on waterbath
they seem to expand and distort and often brittle. So that it looks like only
fibrous tissue is left, the structure is gone. I already had a look on histonet
archive but did not find anything that helps, therefore I hope that someone has
an idea what can be wrong. I have to say that we dont have any problems when
we immersion fix the brains with 4% formaldehyde.
The problems we have with rat brains are a bit different, because we get the
brains from another group who perfuse the lung with 4% paraformaldehyde in
Hepes-buffer, pH 7.35. We then postfix over night in the same fixative they
use (when the brain is also perfused) or for 3 days (when brain is not
perfused). After dehydration in tissue processor we have the same cutting
problems like we observe with mouse brain tissue, even worse.
Any suggestions are appreciated!

Thank you very much for your help!!!

Martina Urbanek



Ms. Martina Urbanek
Forschungslabor der
Klin.Abt. fr Neonatologie
neonatal neuroscience research laboratory
Med. University Innsbruck
Innrain 66, 4th floor
A-6020 Innsbruck
Tel. +43 (0)512 504 27755/27765
Fax: +43 (0)512 504 27766
Email: Martina.Urbanek@uibk.ac.at











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

Message: 13
Date: Tue, 17 Oct 2006 12:52:24 -0400
From: "Goodwin, Diana" 
Subject: [Histonet] Phospho-S6 Ribosomal Protein
To: ,
	
Message-ID:
	<80CDD9C3FEEAFD4982B114C4A6DFD00E02CB8235@uphsmbx2.UPHS.PENNHEALTH.PRV>
	
Content-Type: text/plain;	charset="us-ascii"

For those of us 'Netters using this Ab on human tissue, which one and at
what dilution?
 
Thanks!
 
Diana Goodwin
Supervisor, Anatomic Pathology
Pennsylvania Hospital
Preston 655-C
ph. 215-829-6532
pager 215-422-5160
fax 215-829-7564
e-mail goodwind@[pahosp.com
 


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