[Histonet] RE: Autoimmunostainers DAKO vs Ventanna

From:"White, Lori "

I have used both systems and there are pros and cons to each.  It really is dependent on your IHC volume and how you want to use the automation.  If you want to be free of working up antibodies from concentrates to maximize dilution and are willing to pay the expense of prediluted reagents, the Ventana system is probably the better way to go.  Keep in mind that you are  locked into one source for all reagents including buffers and detection.  To me, this is a disadvantage.
If you have a fairly high volume IHC laboratory and experienced staff, I would recommend the open systems.  You have a lot more flexibility for not only primary antibodies but all of your ancillary reagents as well.  One of the features I liked about the Dako was that you were able to see what was happening during the staining run, not only on the instrument but on the computer screen as well.  I also liked the sensor/alarm for dispensing of reagents.
In terms of service, I would say that again both systems have positives and negatives.
My two cents......

Lori White
Charge Technologist
Lakeridge Health Corporation
Oshawa, Ontario Canada

-----Original Message-----
From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] 
Sent: Wednesday, December 01, 2004 7:26 AM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 13, Issue 1

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Today's Topics:

   1. Ammonia water pH (Paula Pierce)
   2. Frozen cell pellet (Susan Q Wells)
   3. RE: Frozen cell pellet (Favara, Cynthia (NIH/NIAID))
   4. Re: Ammonia water pH and other bluing solutions (Johnson, Teri)
   5. gliut/isopropyl alcohol (Molinari, Betsy)
   6. Re: Re: Ammonia water pH and other bluing solutions
      (Jackie M O'Connor)
   7. Re: Re: Ammonia water pH and other bluing solutions
      (Robyn Vazquez)
   8. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James)
   9. Re: Frozen cell pellet (Gayle Callis)
  10. Saturated lithium carbonate bluing  (Gayle Callis)
  11. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James)
  12. Re: Ammonia water pH and other bluing solutions (Johnson, Teri)
  13. Paraffin processing explant cultures (Johnson, Teri)
  14. Re: Re: Ammonia water pH and other bluing solutions (Gudrun Lang)
  15. RE: Paraffin blocks - Dangerous goods? (Luck, Greg D.)
  16. RE: Counterstain for Von Kossa (Fredrik Johansson)
  17. Phosphorylated-met antibodies (Tan, MinHan)
  18. RE: Paraffin processing explant cultures (Elizabeth Chlipala)
  19. CD 133 (nperson211@comcast.net)
  20. RE: Paraffin processing explant cultures (Johnson, Teri)
  21. RE: Paraffin blocks - Dangerous goods? (Robyn Vazquez)
  22. Re: Ammonia water pH (SMITH,REBEKAH FELICIA)
  23. Re: Paraffin blocks - Dangerous goods? (Jennifer MacDonald)
  24. Re: gliut/isopropyl alcohol (RCHIOVETTI@aol.com)
  25. Part time Histology position in San Diego Ca. (James Watson)
  26. RE: Re: Ammonia water pH and other bluing solutions[Sc	anned]
      (Kemlo Rogerson)
  27. RE: gliut/isopropyl alcohol[Scanned] (Kemlo Rogerson)
  28. RE: gliut/isopropyl alcohol (Molinari, Betsy)


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

Message: 1
Date: Tue, 30 Nov 2004 10:17:12 -0800 (PST)
From: Paula Pierce 
Subject: [Histonet] Ammonia water pH
To: histonet@lists.utsouthwestern.edu
Message-ID: <20041130181712.32329.qmail@web50303.mail.yahoo.com>
Content-Type: text/plain; charset=us-ascii

lol, Jackie O'Conner. I like you. I do the same thing.



Paula Pierce, HTL(ASCP)HT

Excalibur Pathology, Inc.
631 N. Broadway
Moore, OK 73160
405-759-3953
contact@excaliburpathology.com
www.excaliburpathology.com

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

Message: 2
Date: Tue, 30 Nov 2004 13:21:51 -0500
From: Susan Q Wells 
Subject: [Histonet] Frozen cell pellet
To: Histonet@lists.utsouthwestern.edu
Message-ID: <41ACBA3F.9030602@bms.com>
Content-Type: text/plain; format=flowed; charset=ISO-8859-1

Does anyone have advise on how to freeze cells that have been spun down 
in a micro centrifuge tube? In the past I've snap frozen
the tube, loosened the cells by flicking the tube and then embedded the 
pellet in OCT. The cells  look OK, but the cells in the middle don't 
look as good as the ones on the perimeter.Any advise would be appreciated.
Thanks,
Sue Wells



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

Message: 3
Date: Tue, 30 Nov 2004 13:38:40 -0500
From: "Favara, Cynthia (NIH/NIAID)" 
Subject: RE: [Histonet] Frozen cell pellet
To: 'Susan Q Wells' ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain

I have done as you have, also spin down add a bit of histogel vortex let
cool then flick out and put in OCT. I like this as I seem to get a more even
distribution of cells.

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317

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-----Original Message-----
From: Susan Q Wells [mailto:susan.wells@bms.com] 
Sent: Tuesday, November 30, 2004 11:22 AM
To: Histonet@lists.utsouthwestern.edu
Subject: [Histonet] Frozen cell pellet

Does anyone have advise on how to freeze cells that have been spun down in a
micro centrifuge tube? In the past I've snap frozen the tube, loosened the
cells by flicking the tube and then embedded the pellet in OCT. The cells
look OK, but the cells in the middle don't look as good as the ones on the
perimeter.Any advise would be appreciated.
Thanks,
Sue Wells

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

Message: 4
Date: Tue, 30 Nov 2004 12:51:22 -0600
From: "Johnson, Teri" 
Subject: [Histonet] Re: Ammonia water pH and other bluing solutions
To: "Histonet" 
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

Way back in the day, Brigati was using PBS on his immunostainer to blue
after the hematoxylin counterstain.  I have since used PBS to blue my
slides instead of ammonium hydroxide/water.  It's cheap, and very easy
on the sections, and I have plenty of it on hand.
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


 


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

Message: 5
Date: Tue, 30 Nov 2004 13:07:43 -0600
From: "Molinari, Betsy" 
Subject: [Histonet] gliut/isopropyl alcohol
To: 
Message-ID: 
Content-Type: text/plain;	charset="us-ascii"

Hi histonetters,
 I received samples that have been fixed   in a   1%glut / 20% isopropyl
alcohol solution for paraffin processing. I have never heard of this
fixative, I know alcoholic formalin but never alcoholic glutaraldehyde.
Are any of you familiar with this fixative?
Thank you.
Betsy Molinari HT (ASCP)
Texas Heart Institute
Cardiovascular Pathology
Houston,TX 77030
832-355-6524


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

Message: 6
Date: Tue, 30 Nov 2004 13:08:41 -0600
From: "Jackie M O'Connor" 
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
	solutions
To: "Johnson, Teri" 
Cc: Histonet ,
	histonet-bounces@lists.utsouthwestern.edu
Message-ID:
	
	
Content-Type: text/plain; charset="us-ascii"

Anyone remember using saturated lithium carbonate?  I could probably use a 
little lithium right now . . . . . 

Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just 
bringing the slides BACK to a neutral pH after treating them with acid 
which makes them purple-ish?  I like the ammonia because it 'seems' to 
make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to 
make them "blah".(Perception is in the eye of the beholder)   Running tap 
water will make the same miracle happen, but it takes longer.






"Johnson, Teri" 
Sent by: histonet-bounces@lists.utsouthwestern.edu
11/30/2004 12:51 PM

 
        To:     "Histonet" 
        cc: 
        Subject:        [Histonet] Re: Ammonia water pH and other bluing solutions


Way back in the day, Brigati was using PBS on his immunostainer to blue
after the hematoxylin counterstain.  I have since used PBS to blue my
slides instead of ammonium hydroxide/water.  It's cheap, and very easy
on the sections, and I have plenty of it on hand.
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


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




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

Message: 7
Date: Tue, 30 Nov 2004 11:38:08 -0800
From: "Robyn Vazquez" 
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
	solutions
To: Jackie.O'Connor@abbott.com,
	histonet-bounces@lists.utsouthwestern.edu,	TJJ@Stowers-Institute.org
Cc: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset=us-ascii

Jackie,
I use a saturated lithium carbonate for my frozen Mohs tissue.  Works
and looks great...
 
Robyn
OHSU

>>> "Jackie M O'Connor"  11/30/2004
11:08:41 AM >>>

Anyone remember using saturated lithium carbonate?  I could probably
use a 
little lithium right now . . . . . 

Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step
just 
bringing the slides BACK to a neutral pH after treating them with acid

which makes them purple-ish?  I like the ammonia because it 'seems' to

make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to

make them "blah".(Perception is in the eye of the beholder)   Running
tap 
water will make the same miracle happen, but it takes longer.






"Johnson, Teri" 
Sent by: histonet-bounces@lists.utsouthwestern.edu
11/30/2004 12:51 PM


        To:     "Histonet" 
        cc: 
        Subject:        [Histonet] Re: Ammonia water pH and other
bluing solutions


Way back in the day, Brigati was using PBS on his immunostainer to
blue
after the hematoxylin counterstain.  I have since used PBS to blue my
slides instead of ammonium hydroxide/water.  It's cheap, and very easy
on the sections, and I have plenty of it on hand.


Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org



_______________________________________________
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



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

Message: 8
Date: Wed, 1 Dec 2004 08:49:29 +1300
From: "Darren James" 
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]
To: "'Kemlo Rogerson'" ,	"Histonet
	\(E-mail\)" 
Message-ID: <000001c4d715$b49168d0$c864a8c0@medica.co.nz>
Content-Type: text/plain;	charset="iso-8859-1"

As another option take a look at the Bond Max manufactured by Vision Bio
Systems
http://www.vision-bio.com/product_bondmax.html

It performs on board dewaxing, heat induced epitope retreival, enzyme
digestion as well as counterstaining.
I have only heard good things about it.

I have used the Dako extensively and although good, it has it's limitations.
At the time we chose the Dako over the Ventana due to the Ventana being a
closed system. If I were in a position to purchase another I would
definitely take a long hard look at the Bond Max.

Thanks and have a great day
Darren James

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo
Rogerson
Sent: Wednesday, 1 December 2004 4:36 a.m.
To: histonet@pathology.swmed.edu
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


WE have both, and I'm told the Ventana is the 'better' of the two options.
As an ICC virgin and one who will have to choose which platform to go with I
am particularly interested in this thread.

I would prefer to use the cheapest 'open' system that is a automated as
possible, is that mutually exclusive?

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194


-----Original Message-----
From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com]
Sent: 30 November 2004 15:16
To: histonet@pathology.swmed.edu
Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


        I am looking for info= rmation from people who have switched from
   Ventanna  to  DAKO  and DAKO to V= entanna.  Why did you switch?  What
   did you like or dislike abo= ut one or the other.

   Rick
   ________________________________________________________________
   Important news about our email communications
   Saint  Barnabas  Health  Care  System has implemented secure messaging
   servic= es.
   To learn more about SBHCS Secure Messaging, go to:
   [1]http://www.zixcorp.c= om/evangelism/sbhcs/
   If  you  need  assistance with retrieving a secure email, please email
   sbhcs= accounts@sbhcs.com or visit
   [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php

References

   1. 3D"http://www.zixcorp.com/evangelism/sbhcs/"
   2.
3D"http://www.zixcorp.com/evangelism/_______________________________________
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------------------------------

Message: 9
Date: Tue, 30 Nov 2004 12:45:41 -0700
From: Gayle Callis 
Subject: Re: [Histonet] Frozen cell pellet
To: Susan Q Wells ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.1.20041130123759.01b57008@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

How many cells you spin down may be critical.  Here is a protocol that 
works for us.

1.  1 to 3 x 10 (to the 7th) cells/ml.
2.  Spin down and rinse three time with PBS.
3.  Add OCT (approx 1 ml or less) to last pellet, mix.
4.  Immerse tube directly into liquid N2.
5.  Pop block out with a sharp rap inside cryostat.

Mount block and section.

The key is to make sure you don't have too many cells and do a good mix 
with OCT in order to disperse any water from buffer and spread out 
cells.  We have sectioned thinner, 4 um rather than 5 to insure uncrowded 
staining.

We build block up with more OCT around end of block so the block face is 
not so tiny and you can manipulate the tiny section.  Chris van der Loos is 
a master at the supertiny section!!

A wider conical end microcentrifuge tube (holds 2 ml or more overall) 
spreads things out a bit more for a broader faced block but a 1.5 ml tube 
will work.
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 10
Date: Tue, 30 Nov 2004 12:58:26 -0700
From: Gayle Callis 
Subject: [Histonet] Saturated lithium carbonate bluing 
To: "Jackie M O'Connor" ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.1.20041130124819.01b66e58@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Yes, have used sat LiCO3, but it had to be decanted carefully into the 
container or the lithium carbonate crystals poured off polarized!!  Bummer 
with congo red amyloid staining to see annoying crystals everywhere.   And 
then the saturated stuff grew some ugly black floating grow-ty stuff, a 
touch of black slime.   Went back to Scotts tap water substitute or Richard 
Allen bluing solution.

An excellent article on the theory of hematoxylin staining is found in J of 
Histotechnology, Susan Meloan and H. Puchtler volume 10, 1987.   The actual 
staining by hematoxylin, nomenclature, and what the dye molecule consists 
of, along with what acid and alkaline solutions does is explained in great 
detail.  After reading this publication, how I do or think about 
hematoxylin staining (either progressive or regressive methods) was 
radically changed.   It is very chemical, but very informative - be 
prepared for some interesting chemistry.

I strongly recommend this publication to all doing H&E staining.


At 12:08 PM 11/30/2004, you wrote:
>Anyone remember using saturated lithium carbonate?  I could probably use a
>little lithium right now . . . . .
>
>Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just
>bringing the slides BACK to a neutral pH after treating them with acid
>which makes them purple-ish?  I like the ammonia because it 'seems' to
>make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to
>make them "blah".(Perception is in the eye of the beholder)   Running tap
>water will make the same miracle happen, but it takes longer.
>
>
>
>

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: 11
Date: Wed, 1 Dec 2004 09:28:05 +1300
From: "Darren James" 
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]
To: "'Bonnie Whitaker'" ,	"Histonet
	\(E-mail\)" 
Message-ID: <000801c4d71b$198aeb80$c864a8c0@medica.co.nz>
Content-Type: text/plain;	charset="us-ascii"

Hi Bonnie,
I don't know about the US and the UK but here in NZ and also Australia they
are being placed at various sites.
I would assume that if there were any pending legal action the hold on
selling would be global.
Thanks
Darren

-----Original Message-----
From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com]
Sent: Wednesday, 1 December 2004 9:01 a.m.
To: darrenj@medica.co.nz
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


I heard that they were held up in litigation, and would be unable to sell
the instruments for at least a year..... Also, it's a closed system (at
least to some degree.)

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Darren James
Sent: Tuesday, November 30, 2004 1:49 PM
To: 'Kemlo Rogerson'; Histonet (E-mail)
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


As another option take a look at the Bond Max manufactured by Vision Bio
Systems http://www.vision-bio.com/product_bondmax.html

It performs on board dewaxing, heat induced epitope retreival, enzyme
digestion as well as counterstaining. I have only heard good things about
it.

I have used the Dako extensively and although good, it has it's limitations.
At the time we chose the Dako over the Ventana due to the Ventana being a
closed system. If I were in a position to purchase another I would
definitely take a long hard look at the Bond Max.

Thanks and have a great day
Darren James

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo
Rogerson
Sent: Wednesday, 1 December 2004 4:36 a.m.
To: histonet@pathology.swmed.edu
Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


WE have both, and I'm told the Ventana is the 'better' of the two options.
As an ICC virgin and one who will have to choose which platform to go with I
am particularly interested in this thread.

I would prefer to use the cheapest 'open' system that is a automated as
possible, is that mutually exclusive?

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194


-----Original Message-----
From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com]
Sent: 30 November 2004 15:16
To: histonet@pathology.swmed.edu
Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned]


        I am looking for info= rmation from people who have switched from
   Ventanna  to  DAKO  and DAKO to V= entanna.  Why did you switch?  What
   did you like or dislike abo= ut one or the other.

   Rick
   ________________________________________________________________
   Important news about our email communications
   Saint  Barnabas  Health  Care  System has implemented secure messaging
   servic= es.
   To learn more about SBHCS Secure Messaging, go to:
   [1]http://www.zixcorp.c= om/evangelism/sbhcs/
   If  you  need  assistance with retrieving a secure email, please email
   sbhcs= accounts@sbhcs.com or visit
   [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php

References

   1. 3D"http://www.zixcorp.com/evangelism/sbhcs/"
   2.
3D"http://www.zixcorp.com/evangelism/_______________________________________
________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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

Message: 12
Date: Tue, 30 Nov 2004 15:09:05 -0600
From: "Johnson, Teri" 
Subject: [Histonet] Re: Ammonia water pH and other bluing solutions
To: "Histonet" 
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

FYI, the pH of the PBS we use is 7.2 but I'm sure any neutral to
slightly alkaline pH would work fine.
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


 


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

Message: 13
Date: Tue, 30 Nov 2004 15:13:43 -0600
From: "Johnson, Teri" 
Subject: [Histonet] Paraffin processing explant cultures
To: "Histonet" 
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

Is anybody doing paraffin processing on explant cultures and can give me
some tips on increasing my chances for success?  The culture plate
inserts are Millicell polycarbonate and my question is whether they can
survive tissue processing and paraffin sectioning.  If not, what are
your recommendations for doing these?
 
Thanks in advance!
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


 


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

Message: 14
Date: Tue, 30 Nov 2004 22:59:33 +0100
From: "Gudrun Lang" 
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
	solutions
To: "Histonetliste" 
Message-ID: <003a01c4d727$e077a3d0$eeeea8c0@server>
Content-Type: text/plain;	charset="iso-8859-1"

Using lithiumcarbonat is the way we do it with staining frozen sections.
In the HE-stainer we have running tapwater. And  the sections are nice.
After haemalaun in special stains we let the slides in tapwater for 3-5 min.

Gudrun Lang


----- Original Message ----- 
From: "Jackie M O'Connor" 
To: "Johnson, Teri" 
Cc: "Histonet" ;

Sent: Tuesday, November 30, 2004 8:08 PM
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions


> Anyone remember using saturated lithium carbonate?  I could probably use a
> little lithium right now . . . . .
>
> Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just
> bringing the slides BACK to a neutral pH after treating them with acid
> which makes them purple-ish?  I like the ammonia because it 'seems' to
> make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to
> make them "blah".(Perception is in the eye of the beholder)   Running tap
> water will make the same miracle happen, but it takes longer.
>
>
>
>
>
>
> "Johnson, Teri" 
> Sent by: histonet-bounces@lists.utsouthwestern.edu
> 11/30/2004 12:51 PM
>
>
>         To:     "Histonet" 
>         cc:
>         Subject:        [Histonet] Re: Ammonia water pH and other bluing
solutions
>
>
> Way back in the day, Brigati was using PBS on his immunostainer to blue
> after the hematoxylin counterstain.  I have since used PBS to blue my
> slides instead of ammonium hydroxide/water.  It's cheap, and very easy
> on the sections, and I have plenty of it on hand.
>
>
> Teri Johnson
> Managing Director Histology Facility
> Stowers Institute for Medical Research
> 1000 E. 50th St.
> Kansas City, Missouri  64110
> tjj@stowers-institute.org
>
>
>
> _______________________________________________
> 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
>




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

Message: 15
Date: Tue, 30 Nov 2004 13:59:19 -0800
From: "Luck, Greg D." 
Subject: RE: [Histonet] Paraffin blocks - Dangerous goods?
To: 'Richard Cartun' ,
	histonet@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain;	charset="iso-8859-1"

Richard,

Would you further define or point me towards a regulatory definition of
"Dangerous Goods".
Thanks, Greg

Greg Luck, BS, HT(ASCP)
Anatomic Pathology Supervisor
Deaconess Medical Center
800 W. 5th Ave
Spokane, WA 99204
Phone 509.473.7077
Fax 509.473.7133
luckg@empirehealth.org



-----Original Message-----
From: Richard Cartun [mailto:Rcartun@harthosp.org]
Sent: Tuesday, November 30, 2004 8:12 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin blocks - Dangerous goods?


I have just been told that paraffin tissue blocks are considered
"Dangerous Goods" and can only be packaged and shipped by individuals
who have received special training.  Would anyone like to comment on
this?  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



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

Message: 16
Date: Tue, 30 Nov 2004 13:59:42 -0800
From: Fredrik Johansson 
Subject: RE: [Histonet] Counterstain for Von Kossa
To: histonet@lists.utsouthwestern.edu
Message-ID: <239E252E-431B-11D9-94FE-000A95F0ED90@u.washington.edu>
Content-Type: text/plain; charset=WINDOWS-1252; format=flowed

Hi

 

Thanks for all the answers about counterstain with Von Kossa.

 

It seemed like most of you preferred nuclear fast red. So I'm going to 
try that.

 

Thanks

 

Fredrik Johansson

Dept. of Pathology

UW, Seattle




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

Message: 17
Date: Tue, 30 Nov 2004 17:02:35 -0500
From: "Tan, MinHan" 
Subject: [Histonet] Phosphorylated-met antibodies
To: 
Message-ID: 
Content-Type: text/plain;	charset="us-ascii"

Good afternoon,

We are thinking of staining for phosphorylated-met; while there are a
number of commercial antibodies out there, it seems that they have not
been tested out on paraffin embedded tissue yet.

If anyone has tried staining for this antigen, I'd appreciate any
pointers you have.

Min-Han
This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential information.  Any unauthorized review, use, disclosure or distribution is prohibited.  If you are not the intended recipient(s) please contact the sender by reply email and destroy all copies of the original message.  Thank you.



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

Message: 18
Date: Tue, 30 Nov 2004 15:02:30 -0700
From: "Elizabeth Chlipala" 
Subject: RE: [Histonet] Paraffin processing explant cultures
To: "'Johnson, Teri'" ,	"'Histonet'"
	
Message-ID: <000001c4d728$49c9c480$76d48a80@AMY>
Content-Type: text/plain;	charset="us-ascii"

Teri

I have processed explant cultures with the Millipore MillicellT CM which
is a polytetrafluoroethylene based polymer.  I process these as any
other tissue.  Millipore has a great web page in which you can ask
questions.  I would give them a call about the particular filter that
you are using. 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson,
Teri
Sent: Tuesday, November 30, 2004 2:14 PM
To: Histonet
Subject: [Histonet] Paraffin processing explant cultures

Is anybody doing paraffin processing on explant cultures and can give me
some tips on increasing my chances for success?  The culture plate
inserts are Millicell polycarbonate and my question is whether they can
survive tissue processing and paraffin sectioning.  If not, what are
your recommendations for doing these?
 
Thanks in advance!
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


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




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

Message: 19
Date: Tue, 30 Nov 2004 22:24:50 +0000
From: nperson211@comcast.net
Subject: [Histonet] CD 133
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<113020042224.12979.41ACF331000E7125000032B32207001641CECECD02019C9D0A9F02@comcast.net>
	
Content-Type: text/plain

Netters,
Is anyone using an antibody against CD 133 on FFPE tissues?  If so, what have you used for a positive control, that could also provide cells to use as a positive control for Flow?  
I would really appreciate any information anyone would be willing to share.
Thanks,
Nancy Lemke
Hermelin Brain Tumor Center
Henry Ford Hospital
Detroit

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

Message: 20
Date: Tue, 30 Nov 2004 17:01:52 -0600
From: "Johnson, Teri" 
Subject: RE: [Histonet] Paraffin processing explant cultures
To: "Histonet" 
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

Liz is a genius.  I called Millipore and their customer service guy,
Rich, told me I could paraffin process and section these with no
problems.  Why didn't *I* think of doing this?  Sheesh...

Teri


-----Original Message-----
From: Elizabeth Chlipala [mailto:liz@premierlab.com] 
Sent: Tuesday, November 30, 2004 4:03 PM
To: Johnson, Teri; 'Histonet'
Subject: RE: [Histonet] Paraffin processing explant cultures


Teri

I have processed explant cultures with the Millipore MillicellT CM which
is a polytetrafluoroethylene based polymer.  I process these as any
other tissue.  Millipore has a great web page in which you can ask
questions.  I would give them a call about the particular filter that
you are using. 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz@premierlab.com
www.premierlab.com
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson,
Teri
Sent: Tuesday, November 30, 2004 2:14 PM
To: Histonet
Subject: [Histonet] Paraffin processing explant cultures

Is anybody doing paraffin processing on explant cultures and can give me
some tips on increasing my chances for success?  The culture plate
inserts are Millicell polycarbonate and my question is whether they can
survive tissue processing and paraffin sectioning.  If not, what are
your recommendations for doing these?
 
Thanks in advance!
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


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




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

Message: 21
Date: Tue, 30 Nov 2004 15:29:32 -0800
From: "Robyn Vazquez" 
Subject: RE: [Histonet] Paraffin blocks - Dangerous goods?
To: LuckG@empirehealth.org, Rcartun@harthosp.org,
	histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset=us-ascii

Richard,
They are a regulated class 6.2, it is a substance known or reasonalbly
expected to contain pathogens.  If you ship dangerous goods you will
need a shipper's declaration which is required.
 
Robyn
OHSU 

>>> "Luck, Greg D."  11/30/2004 1:59:19 PM >>>

Richard,

Would you further define or point me towards a regulatory definition
of
"Dangerous Goods".
Thanks, Greg

Greg Luck, BS, HT(ASCP)
Anatomic Pathology Supervisor
Deaconess Medical Center
800 W. 5th Ave
Spokane, WA 99204
Phone 509.473.7077
Fax 509.473.7133
luckg@empirehealth.org



-----Original Message-----
From: Richard Cartun [mailto:Rcartun@harthosp.org]
Sent: Tuesday, November 30, 2004 8:12 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin blocks - Dangerous goods?


I have just been told that paraffin tissue blocks are considered
"Dangerous Goods" and can only be packaged and shipped by individuals
who have received special training.  Would anyone like to comment on
this?  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

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



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

Message: 22
Date: Tue, 30 Nov 2004 19:13:50 -0500 (EST)
From: "SMITH,REBEKAH FELICIA" 
Subject: Re: [Histonet] Ammonia water pH
To: "Jackie M O'Connor" ,	Tim Webster
	
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
	<934672373.1101860030245.JavaMail.osg@osgjas02.cns.ufl.edu>
Content-Type: text/plain; format=flowed; charset=us-ascii

I agree with Jackie, I use a Coplin jar with distilled water and 
about 3 drops of 30% ammonium hydroxide, and then rinse in tap 
water twice. i've never had problems and it produces a rather 
pretty blue for me.
                                   Rebekah Smith
On Tue Nov 30 10:35:55 EST 2004, Jackie M O'Connor 
 wrote:

> Maybe I'm being extremely simple-minded - but I just fill up a 
> 200 ml staining jar with tap H20, and add a couple-three drops of 
> 28%.  When it doesn't smell like ammonia anymore, I make new 
> stuff.  How's that grab ya for QA? "Tim Webster" 
> 
> Sent by: histonet-bounces@lists.utsouthwestern.edu
> 11/30/2004 09:30 AM
> 
>          To:     
>         cc:         Subject:        [Histonet] Ammonia water pH
> 
> 
> Hi All,
> 
> I'm curious to know what people's procedures are for making and 
> using ammonia water as a bluing agent in regressive H&E's.
> Currently, our methodology is to make the ammonia water up daily 
> from a 28% stock bottle, and the pH is about 11.  My feeling is 
> that 11 is way to high and that 9-10 is plenty alkaline to blue 
> sections adequately. Additionally, I'm concerned about strong 
> alkaline solution increasing the likelihood of sections washing 
> off or wrinkling.  Sheehan & Hrapchak call for a "weak" alkaline 
> solution in "Theory and Practice of Histotechnology". Also, I can 
> see no reason not to make a working solution weekly/monthly and 
> simply empty and refill the Ammonia water station.  The pH 
> doesn't appear to drop off over time.
> 
> Thanks for your time, and have a great week! Tim
> 
> Tim Webster
> Histology Specialist
> Northwestern Medical Center
> 133 Fairfield Street,
> St Albans, VT 05478
> (802) 524-1070 (x4349)
> twebster@nmcinc.org
> 
>>                                                Statement of
> Confidentiality
>>                                                This message and 
>> any
> attachments are from NMC and intended only for the addressee(s).  
> The information contained may include privileged or other 
> confidential information.  Unauthorized review, forwarding, 
> printing, copying or distribution is strictly prohibited.
>>                                                If you should 
>> receive
> this message in error, please delete it and notify the sender.  
> Thank you.
>> 
>> 
> _______________________________________________
> 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
> 
> 



--
SMITH,REBEKAH FELICIA
"You are a child of the universe, no less than the trees and the 
stars
You have a right to be here and whether or not it is clear to you, 
no doubt the universe is unfolding as it should. Therefore be at 
peace with G-d, whatever you conceive Him to be. And whatever your 
labors and aspirations,in the noisy confusion of life, keep peace 
in your soul.-Max Ehrmann,"Desiderata"




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

Message: 23
Date: Tue, 30 Nov 2004 16:17:46 -0800
From: Jennifer MacDonald 
Subject: Re: [Histonet] Paraffin blocks - Dangerous goods?
To: "Richard Cartun" 
Cc: histonet@lists.utsouthwestern.edu,
	histonet-bounces@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain; charset="US-ASCII"

Advance for Medical Laboratory Professionals had a good article in the 
November 1, 2004 issue.  It is under the Q&A section, titled:  Standards 
for Shipping Infectious Agents.  Check www.advanceweb.com to check the 
archives.  If you can't find it, I can fax you a copy.  There is also an 
old article titled "Safe Specimen Packaging Begins with You.  It is from 
the November 12, 2001 issue.

Jennifer MacDonald
Mt. San Antonio College





"Richard Cartun"  
Sent by: histonet-bounces@lists.utsouthwestern.edu
11/30/2004 08:12 AM

To

cc

Subject
[Histonet] Paraffin blocks - Dangerous goods?






I have just been told that paraffin tissue blocks are considered
"Dangerous Goods" and can only be packaged and shipped by individuals
who have received special training.  Would anyone like to comment on
this?  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



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

Message: 24
Date: Tue, 30 Nov 2004 19:26:06 EST
From: RCHIOVETTI@aol.com
Subject: Re: [Histonet] gliut/isopropyl alcohol
To: BMolinari@heart.thi.tmc.edu, histonet@lists.utsouthwestern.edu
Message-ID: <1cc.2d22ffb1.2ede699e@aol.com>
Content-Type: text/plain; charset="US-ASCII"

In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, 
BMolinari@heart.thi.tmc.edu writes:
I received samples that have been fixed   in a   1%glut / 20% isopropyl
alcohol solution for paraffin processing. I have never heard of this
fixative, I know alcoholic formalin but never alcoholic glutaraldehyde.
Are any of you familiar with this fixative?
Thank you.
Betsy Molinari HT (ASCP)
Hi Betsy,

Is the specimen perhaps a heart valve made from pig tissue??

The only time I've heard of alcoholic glutaraldehyde is for pretreating pig 
valves before they're implanted in humans.  There's some kind of relationship 
between alcohol and glut, and preventing calcification of the valve.  Can't 
remember the details right now, but you have to treat the valve with one of those 
reagents before using the other.

Just a thought...

Cheers,

Bob Chiovetti


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

Message: 25
Date: Tue, 30 Nov 2004 17:28:20 -0800
From: "James Watson" 
Subject: [Histonet] Part time Histology position in San Diego Ca.
To: 
Message-ID:
	
Content-Type: text/plain;	charset="US-ASCII"

GNF in San Diego has a part time position available.  Hours are
flexible, 20 hours per week.  Please e-mail resume or call me at the
number below.

The position requires ASCP HT certification.  BS, AA degree, H.S. with
completion of certified Histology Program with a minimum of 3 years of
experience, or equivalent training with a minimum of 6 years of
experience.  Must be experienced with frozen sectioning, tissue
processing and embedding, paraffin sectioning, H&E and special staining,
and immunohistochemistry.  Animal tissue trimming and identification is
preferred.


James Watson HT, ASCP
Facilities Manager of Histology
GNF, Genomics Institute of the Novartis Research Foundation
Room C015 
858-332-4647
jwatson@gnf.org 



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

Message: 26
Date: Wed, 1 Dec 2004 08:18:41 -0000 
From: Kemlo Rogerson 
Subject: RE: [Histonet] Re: Ammonia water pH and other bluing
	solutions[Sc	anned]
To: 'Jackie M O'Connor' , "Johnson, Teri"
	
Cc: Histonet ,
	histonet-bounces@lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B54C@bhrv-nt-11.bhrv.nwest.nhs.uk>
	
Content-Type: text/plain

I think you are wrong, so I'll correct you, as you requested.

I thought the 'blue' state of haematin was, like litmus, the effect an
alkaline condition has on the siderphilic dye. Red, like litmus, denoted
acidic conditions. Damned if I know what colour (with the 'u') haematin goes
when neutral (bluey/ red?)

Running tap water takes longer, I concede, but good things are worth waiting
for and it's easier to make up; we rush too much I find. TIP: Never use
London water, been through too many kidneys!

-----Original Message-----
From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] 
Sent: 30 November 2004 19:09
To: Johnson, Teri
Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing
solutions[Scanned]

Anyone remember using saturated lithium carbonate?  I could probably use a 
little lithium right now . . . . . 

Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just 
bringing the slides BACK to a neutral pH after treating them with acid 
which makes them purple-ish?  I like the ammonia because it 'seems' to 
make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to 
make them "blah".(Perception is in the eye of the beholder)   Running tap 
water will make the same miracle happen, but it takes longer.






"Johnson, Teri" 
Sent by: histonet-bounces@lists.utsouthwestern.edu
11/30/2004 12:51 PM

 
        To:     "Histonet" 
        cc: 
        Subject:        [Histonet] Re: Ammonia water pH and other bluing
solutions


Way back in the day, Brigati was using PBS on his immunostainer to blue
after the hematoxylin counterstain.  I have since used PBS to blue my
slides instead of ammonium hydroxide/water.  It's cheap, and very easy
on the sections, and I have plenty of it on hand.
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


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


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

Message: 27
Date: Wed, 1 Dec 2004 08:25:23 -0000 
From: Kemlo Rogerson 
Subject: RE: [Histonet] gliut/isopropyl alcohol[Scanned]
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<1030B679AD69D6119C3F00080210DD9D01B0B54D@bhrv-nt-11.bhrv.nwest.nhs.uk>
	
Content-Type: text/plain

Expect the tissue is rather brittle, used to be used for electromicroscopy I
think. Wonder if you could secondary fix? Can't aldehyde/ alcohol fixation
be 'washed out' then the tissue refixed in something more conventional? 

-----Original Message-----
From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] 
Sent: 30 November 2004 19:08
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] gliut/isopropyl alcohol[Scanned]

Hi histonetters,
 I received samples that have been fixed   in a   1%glut / 20% isopropyl
alcohol solution for paraffin processing. I have never heard of this
fixative, I know alcoholic formalin but never alcoholic glutaraldehyde.
Are any of you familiar with this fixative?
Thank you.
Betsy Molinari HT (ASCP)
Texas Heart Institute
Cardiovascular Pathology
Houston,TX 77030
832-355-6524
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 28
Date: Wed, 1 Dec 2004 06:18:48 -0600
From: "Molinari, Betsy" 
Subject: RE: [Histonet] gliut/isopropyl alcohol
To: ,	
Message-ID: 
Content-Type: text/plain;	charset="us-ascii"

Hi Bob,
Thanks for your input. I may ask a couple of the surgery staff here if
they are familiar with that technique. A point of interest, these are
valves. I loaded them last night starting in 70% ETOH, I am curious to
see how they turned out and how much effect it will have on the B&B and
PTAH I will be running on them
Thanks again,
Betsy Molinari  HT (ASCP)
Texas Heart Institute
Cardiovascular Pathology
Houston, TX 77030
832-355-6524
 
-----Original Message-----
From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] 
Sent: Tuesday, November 30, 2004 6:26 PM
To: Molinari, Betsy; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] gliut/isopropyl alcohol
 
In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time,
BMolinari@heart.thi.tmc.edu writes:
	I received samples that have been fixed   in a   1%glut / 20%
isopropyl
	alcohol solution for paraffin processing. I have never heard of
this
	fixative, I know alcoholic formalin but never alcoholic
glutaraldehyde.
	Are any of you familiar with this fixative?
	Thank you.
	Betsy Molinari HT (ASCP)
Hi Betsy,
 
Is the specimen perhaps a heart valve made from pig tissue??
 
The only time I've heard of alcoholic glutaraldehyde is for pretreating
pig valves before they're implanted in humans.  There's some kind of
relationship between alcohol and glut, and preventing calcification of
the valve.  Can't remember the details right now, but you have to treat
the valve with one of those reagents before using the other.
 
Just a thought...
 
Cheers,
 
Bob Chiovetti


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

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

End of Histonet Digest, Vol 13, Issue 1
***************************************

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