[Histonet] immunostaining for sca-1

From:Melissa Mazan



Hi all, we're trying to stain simultaneously for sca-1 and SPC in mouse 
lung tissue - I'm unable to get the sca-1 to work (using a rat 
anti-mouse clone from BD) using immunofluorescence - so am trying the 
Vector ABC system.  Unfortunately, to get the sca-1 to work, I have to 
leave out triton-X - without the Triton-X the SPC seems not to be 
working.  Does anyone have advice for trying to simultaneously stain for 
an intracytoplasmic protein and a membrane protein at the same time? Is 
there a better fixative than 4% formaldehyde ( we leave the tissues in 
formaldehyde from 6hours to no more than 48 hours).  Many thanks - 
Melissa Mazan



Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor and Director of Equine Sports Medicine
Department of Clinical Sciences
Tufts Cumming School of Veterinary Medicine
200 Westborough Road
North Grafton, MA 01536
Tel:508-839-5395
Fax:508-839-7922
email: melissa.mazan@tufts.edu

histonet-request@lists.utsouthwestern.edu wrote:
> Send Histonet mailing list submissions to
> 	histonet@lists.utsouthwestern.edu
>
> To subscribe or unsubscribe via the World Wide Web, visit
> 	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> or, via email, send a message with subject or body 'help' to
> 	histonet-request@lists.utsouthwestern.edu
>
> You can reach the person managing the list at
> 	histonet-owner@lists.utsouthwestern.edu
>
> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
>
>
> Today's Topics:
>
>    1. 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/
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
> ------------------------------
>
> 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 4.5 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
>
> _________________________________________________________________
> 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
>
>
>
>
>  		
> ---------------------------------
>  All-new Yahoo! Mail - Fire up a more powerful email and get things done faster. 
>
> ------------------------------
>
> 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
>
>
>                                         
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> *****************************************************************
> This email, including any attachments sent with it, is
> confidential and for the sole use of the intended recipient(s).
> This confidentiality is not waived or lost, if you receive it and
> you are not the intended recipient(s), or if it is transmitted/
> received in error.
>
> Any unauthorised use, alteration, disclosure, distribution or
> review of this email is strictly prohibited.  The information
> contained in this email, including any attachment sent with
> it, may be subject to a statutory duty of confidentiality if it
> relates to health service matters.
>
> If you are not the intended recipient(s), or if you have
> received this email in error, you are asked to immediately
> notify the sender by telephone collect on Australia
> +61 1800 198 175 or by return email.  You should also
> delete this email, and any copies, from your computer
> system network and destroy any hard copies produced.
>
> If not an intended recipient of this email, you must not copy,
> distribute or take any action(s) that relies on it; any form of
> disclosure, modification, distribution and/or publication of this
> email is also prohibited.
>
> Although Queensland Health takes all reasonable steps to
> ensure this email does not contain malicious software,
> Queensland Health does not accept responsibility for the
> consequences if any person's computer inadvertently suffers
> any disruption to services, loss of information, harm or is
> infected with a virus, other malicious computer programme or
> code that may occur as a consequence of receiving this
> email.
>
> Unless stated otherwise, this email represents only the views
> of the sender and not the views of the Queensland Government.
> ****************************************************************
>
>
>
>
> ------------------------------
>
> 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. 
>
> -----------------------------------------
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>  		
> ---------------------------------
> Get your email and more, right on the  new Yahoo.com 
>
> ------------------------------
>
> 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
>
>
>
>
>
>
> -------------- next part --------------
> BEGIN:VCARD
> VERSION:2.1
> X-GWTYPE:USER
> FN:Peter Rippstein
> EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca
> N:Rippstein;Peter
> END:VCARD
>
> BEGIN:VCARD
> VERSION:2.1
> X-GWTYPE:USER
> FN:Peter Rippstein
> EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca
> N:Rippstein;Peter
> END:VCARD
>
> BEGIN:VCARD
> VERSION:2.1
> X-GWTYPE:USER
> FN:Peter Rippstein
> EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca
> N:Rippstein;Peter
> END:VCARD
>
> BEGIN:VCARD
> VERSION:2.1
> X-GWTYPE:USER
> FN:Peter Rippstein
> EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca
> N:Rippstein;Peter
> END:VCARD
>
>
> ------------------------------
>
> 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
>  
>
>
> The information contained in this e-mail message is intended only for the personal and confidential use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you are hereby notified that you have received this document in error and that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately by e-mail, and delete the original message.
>
> ------------------------------
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> End of Histonet Digest, Vol 35, Issue 28
> ****************************************
>   

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




<< Previous Message | Next Message >>