[Histonet] Re: Precipitate in Processor
Adrienne Aperghis Kavanagh
aaperghis <@t> uspath.com
Mon Aug 9 12:14:02 CDT 2010
Thank you all for your quick and helpful responses! I find it strange that we haven't encountered this problem until today, since we have been using the same NBF and alcohols...
We usually do the processor hot water flush weekly, on the 1st four stations. I will flush the 1st seven stations and will do it twice to be sure we get out any leftover precipitate.
In addition to the flush, I will change the 70% alcohol daily until we receive in another formalin.
Thanks again all!
Adrienne Aperghis Kavanagh
US PATH
30 W. Century Road
Suite 255
Paramus NJ 07652
----- Original Message -----
From: histonet-request <@t> lists.utsouthwestern.edu
To: histonet <@t> lists.utsouthwestern.edu
Sent: Monday, August 9, 2010 1:03:43 PM
Subject: Histonet Digest, Vol 81, Issue 9
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Today's Topics:
1.
CA4DF32ED505D94BB55E95487D8E984104FD48 <@t> DOAISD5205.state.mt.ads (Andrew
Burgeson) 2. Re tunel staining (Steven Weston)
3. Re: [Special Stains] Masson's Trichrome (Sherwood, Margaret )
4. RE: friable or crumbly O.C.T. (Della Speranza, Vinnie)
5. Re: friable or crumbly O.C.T. (jsjurczak <@t> comcast.net)
6. Anti-IDO (indolamine 2,3-dioxygenase) (Mark Tarango)
7. Re: Re: [Special Stains] Masson's Trichrome (Rena Fail)
8. RELIA Histology Job Alert 8-09-10 Histotech needed in
Charlotte, NC (Pam Barker)
9. RE: friable or crumbly O.C.T. (Bill B.)
10. Precipitate in Processor (Adrienne Aperghis Kavanagh)
11. Re: Precipitate in Processor (Brandi Higgins)
12. RELIA Solutions Hot Histology Job Alert!! Histotech needed in
Indianapolis area. (Pam Barker)
13. AW: [Histonet] Re: [Special Stains] Masson's Trichrome
(Gudrun Lang)
14. RE: Precipitate in Processor (Jon St.Onge)
15. Re: Precipitate in Processor (Drew Meyer)
16. Re: Stability of Special Stains (Sherwood, Margaret )
----------------------------------------------------------------------
Message: 1
Date: Sun, 08 Aug 2010 13:10:54 -0400
From: "Andrew Burgeson" <napoli <@t> siscom.net>
Subject: [Histonet]
CA4DF32ED505D94BB55E95487D8E984104FD48 <@t> DOAISD5205.state.mt.ads
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <4c5ee51e.73.527e.1030580047 <@t> siscom.net>
Content-Type: text/plain; charset="iso-8859-1"
I am wondering how much agitation is required in order to
achieve the clearing of paraffin from the tissue and slide.
IHC systems obviously use adhesive or + slides, aiding in
tissue adherence. Too much agitation might take take tissue
off. And what about nail fragments or hard tissues in
general? Will they survive? I have an article and info
regarding this type of deparaffinization and will try to get
it into an e-mail.
My sense is that it could very well be less efficient and
time consuming and that depending on how it is done, could
yield very different results.
I still think that "there's nothing like xylene!!!" But
obviously soap and water doesnt hurt your liver....
Interesting method I would like to hear more about people's
experiences.
------------------------------
Message: 2
Date: Mon, 9 Aug 2010 14:32:52 +1000
From: Steven Weston <Steven.Weston <@t> utas.edu.au>
Subject: [Histonet] Re tunel staining
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu> Message-ID:
<C885C214.2618%steven.weston <@t> utas.edu.au> Content-Type: text/plain;
charset="big5"
I recently had an enquiry from one of our post docs who was looking at
TUNEL staining and came across this reference in biochemica no 4
(1997)titled ��Fixation of Tissue Sections for
TUNEL Combined with Staining for
Thymic Epithelial Cell Marker��
You should be able to google it and find the article.
Basically it said that after fixation with PFA to improve staining you
need to treat with triton and sodium citrate in the cold to expose the
antigen again.
Below is the suggested protocol for thin cryostat sections. I��m sure it
could be modified for thick sections.
1. Cut 4 �gm thin cryosections and mount on
polylysine-coated glass slides.
2. Air dry overnight at room temperature and
freeze foil- wrapped slides at �V20�XC until use.
3. Apply frozen glass slides directly into a container
with 1% buffered paraformaldehyde for
30 min at RT.
4. Rinse swiftly in PBS and immerse in a solution
of 1% Triton X100 (v/v) and 1% sodium
citrate (w/v) for 2 min at 4�XC.
5. Wash in PBS and incubate with 50 �gl TUNEL
reaction mixture (TdT solution with dUTP-FITC
solution, 1+9). Convert to enzyme label if
desired. 6. Wash with PBS and block with dilution of
adequate normal serum.
10. Proceed with desired immunohistochemical
labeling.
Regards
Steve Weston
Senior technical officer
Menzies Research Institute
Hobart Tasmania
------------------------------
Message: 3
Date: Mon, 9 Aug 2010 10:32:48 -0400
From: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>
Subject: [Histonet] Re: [Special Stains] Masson's Trichrome
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9703E24532 <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
To all:
We keep having the same discussion re: special stains. How long can you
use them before discarding? I am specifically referring to the stains
used in
Masson's Trichrome.
We are a research lab and, therefore, don't run the volume that most
labs do.
We have found, for instance, that we can use the Weigert's Hematoxylin
(A&B) more than once, with filtering (even though it is stated it should
be made
fresh). We also use Bouin's Fix, Biebrich Scarlet and Aniline Blue more
than once. Is there a "set rule" when we should discard these solutions
(i.e. after
5X use or 1 month)?
I would appreciate some feedback from the experts!
Thanks!
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
The information in this e-mail is intended only for the person to whom
it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners
Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you
in error
but does not contain patient information, please contact the sender and
properly dispose of the e-mail.
------------------------------
Message: 4
Date: Mon, 9 Aug 2010 10:40:03 -0400
From: "Della Speranza, Vinnie" <dellav <@t> musc.edu>
Subject: RE: [Histonet] friable or crumbly O.C.T.
To: "'Bruce W Brodersen'" <bbroders <@t> unlnotes.unl.edu>,
"histonet <@t> pathology.swmed.edu" <histonet <@t> pathology.swmed.edu>
Message-ID:
<E58D1CD977E29C46A167806513B045779AAC134901 <@t> EVS5.clinlan.local>
Content-Type: text/plain; charset="us-ascii"
I'm guessing that liquid nitrogen or dry ice temperature is too cold for
sectioning OCT.
OCT cuts well down to about -25 degrees C.
Liquid Nitrogen is about -160 degrees C. I believe dry ice is in the
same temp range at LN2
You will want to give the OCT blocks the opportunity to "warm up" to
cryostat temperature before attempting to section them. Leave your
frozen blocks in the cryostat for 30-60 minutes before sectioning to
allow them to come to optimum temperature
Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue Suite 309
Charleston, South Carolina 29425
Tel: (843) 792-6353
Fax: (843) 792-8974
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bruce W
Brodersen
Sent: Friday, August 06, 2010 2:26 PM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] friable or crumbly O.C.T.
Anyone have an explanation as to why OCT would be friable or crumbly for
sectioning? Here's how it was used.
Thanks.
"We held the plastic 'tray' with the tissue in the compound just over
the liquid nitro for 30sec-1min, until it was opaque and white (frozen)
and then dipped the tray into the liquid nitro for 20-30sec., placed in
small bags and then into a cooler with dry ice until shipping."
Bruce W. Brodersen, DVM, PhD
University of Nebraska Veterinary Diagnostic Center
1900 N. 42nd Street
Lincoln, NE 68583-0907
voice (402) 472-1434
FAX (402 472-3094_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 5
Date: Mon, 9 Aug 2010 14:49:11 +0000 (UTC)
From: jsjurczak <@t> comcast.net
Subject: Re: [Histonet] friable or crumbly O.C.T.
To: "Della Speranza, Vinnie" <dellav <@t> musc.edu>
Cc: histonet <@t> pathology.swmed.edu
Message-ID:
<2100844269.41947.1281365351046.JavaMail.root <@t> sz0094a.emeryville.ca.mail.comcast.net>
Content-Type: text/plain; charset=utf-8
There is an OCT for lower temperatures.
----- Original Message -----
From: "Della Speranza, Vinnie" <dellav <@t> musc.edu>
To: "Bruce W Brodersen" <bbroders <@t> unlnotes.unl.edu>,
histonet <@t> pathology.swmed.edu
Sent: Monday, August 9, 2010 9:40:03 AM
Subject: RE: [Histonet] friable or crumbly O.C.T.
I'm guessing that liquid nitrogen or dry ice temperature is too cold for
sectioning OCT.
OCT cuts well down to about -25 degrees C.
Liquid Nitrogen is about -160 degrees C. I believe dry ice is in the
same temp range at LN2
You will want to give the OCT blocks the opportunity to "warm up" to
cryostat temperature before attempting to section them. Leave your
frozen blocks in the cryostat for 30-60 minutes before sectioning to
allow them to come to optimum temperature
Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue ��Suite 309
Charleston, South Carolina 29425
Tel: (843) 792-6353
Fax: (843) 792-8974
��
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bruce W
Brodersen
Sent: Friday, August 06, 2010 2:26 PM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] friable or crumbly O.C.T.
Anyone have an explanation as to why OCT would be friable or crumbly for
sectioning? ��Here's how it was used.
Thanks.
"We held the plastic 'tray' with the tissue in the compound just over
the liquid nitro for 30sec-1min, until it was opaque and white (frozen)
and then dipped the tray into the liquid nitro for 20-30sec., placed in
small bags and then into a cooler with dry ice until shipping."
Bruce W. Brodersen, DVM, PhD
University of Nebraska Veterinary Diagnostic Center
1900 N. 42nd Street
Lincoln, NE ��68583-0907
voice (402) 472-1434
FAX (402 472-3094_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________ Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Mon, 9 Aug 2010 07:53:48 -0700
From: Mark Tarango <marktarango <@t> gmail.com>
Subject: [Histonet] Anti-IDO (indolamine 2,3-dioxygenase)
To: "histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<AANLkTikdmxscS6Eefwz8AnpyGW1VDPovg0F=-W2-v51n <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi Histonet,
Does anyone use an Anti-IDO on FFPE tissues? Would you please let me
know where you purchase it?
Thank you,
Mark Tarango
------------------------------
Message: 7
Date: Mon, 9 Aug 2010 08:24:01 -0700 (PDT)
From: Rena Fail <renafail <@t> bellsouth.net>
Subject: Re: [Histonet] Re: [Special Stains] Masson's Trichrome
To: "Sherwood, Margaret" <MSHERWOOD <@t> PARTNERS.ORG>,
histonet <@t> lists.utsouthwestern.edu Message-ID:
<426670.14254.qm <@t> web180808.mail.gq1.yahoo.com> Content-Type: text/plain;
charset=iso-8859-1
It depends on the number of slides as well as time on the shelf and a
little common sense.Your stock solutions have a longer shelf life
than�your working. �I
worked in a lab that perfomed a large number of Masson's by hand( until
purchasing the Artisan)� We changed the solutions once a week to insure
consistent staining.� If you perform these stains only twice a month,
your results will be more consistant if you make the Weigert's fresh�
and watch the
level of your aniline blue, evaporation will make it more concentrated.�
On the
other hand a large volume of slides will weaken the�working solutions.
following is a link that lists some guidelines for storing stock
solutions
Rena Fail
www.urmc.rochester.edu/.../StainsManual/STABILITYOFSPECIALSTAININGSOLUTIONS.html?...-
Cached
----- Original Message ----
From: "Sherwood, Margaret" <MSHERWOOD <@t> PARTNERS.ORG>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Mon, August 9, 2010 10:32:48 AM
Subject: [Histonet] Re: [Special Stains] Masson's Trichrome
To all:
We keep having the same discussion re:� special stains.� How long can
you use
them before discarding?� I am specifically referring to the stains used
in Masson's Trichrome.
We are a research lab and, therefore, don't run the volume that most
labs do.
We have found, for instance, that we can use the Weigert's Hematoxylin
(A&B) more than once, with filtering (even though it is stated it should
be made
fresh).� We also use Bouin's Fix, Biebrich Scarlet and Aniline Blue more
than once.� Is there a "set rule" when we should discard these solutions
(i.e. after
5X use or 1 month)?
I would appreciate some feedback from the experts!
Thanks!
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
The information in this e-mail is intended only for the person to whom
it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners
Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you
in error
but does not contain patient information, please contact the sender and
properly dispose of the e-mail.
_______________________________________________ Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Mon, 9 Aug 2010 11:50:21 -0400
From: "Pam Barker" <relia1 <@t> earthlink.net>
Subject: [Histonet] RELIA Histology Job Alert 8-09-10 Histotech needed
in Charlotte, NC
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <E1OiUcG-0004lN-Kk <@t> elasmtp-curtail.atl.sa.earthlink.net>
Content-Type: text/plain; charset="iso-8859-1"
Hi Histonetters!!
I hope everyone had a great weekend. I have an exciting new opportunity
to tell you about. I am working with a premier client located Charlotte,
NC. This is a part time (32 hours per week) permanent position in a full
service histology lab. My client is looking for an ASCP certified
histotech with at least 2 years of histology experience must be able to
meet CLIA
requirements to do grossing. (My client does mainly simple grossing,
mainly biopsies and excisions). My client offers excellent pay and
benefits. If you or anyone you know would like more information please
contact me at
866-607-3542 or relia1 <@t> earthlink.net
Have a Great Day!!
Thank You!
Pam Barker
President RELIA
Specialists in Allied Healthcare Recruiting
5703 Red Bug Lake Road #330
Winter Springs, FL 32708-4969
Phone: (407)657-2027
Cell: (407)353-5070
FAX: (407)678-2788
E-mail: relia1 <@t> earthlink.net
www.facebook.com search Pam Barker RELIA
www.linkedin.com/reliasolutions www.myspace.com/pamatrelia
www.twitter.com/pamatrelia
------------------------------
Message: 9
Date: Mon, 9 Aug 2010 10:55:40 -0500
From: "Bill B." <bill501 <@t> mindspring.com>
Subject: RE: [Histonet] friable or crumbly O.C.T.
To: "histonet <@t> pathology.swmed.edu" <histonet <@t> pathology.swmed.edu>
Message-ID: <p06240801c885d3bf1ca7@[4.244.63.227]>
Content-Type: text/plain; charset="us-ascii"
I will 2nd this.
When I did neuropathology at a major institution, we froze all frozen
sections in an isopentane slurry cooled with LN2. We waited for the OCT
to warm to cryostate temps before cutting. If there was time pressure
from the surgeons, I used my thumb to warm more quickly, until sections
stopped falling apart. We got minimum freeze artifact with this method.
For research we used homogenized brain (brain paste) instead of OCT
which gave better sections as there was not a change in physical
properties you get with OCT vs brain.
Bill Blank, MD
At 10:40 AM -0400 8/9/10, Della Speranza, Vinnie wrote:
>I'm guessing that liquid nitrogen or dry ice temperature is too cold
>for sectioning OCT.
>OCT cuts well down to about -25 degrees C.
>Liquid Nitrogen is about -160 degrees C. I believe dry ice is in the
>same temp range at LN2
>
>You will want to give the OCT blocks the opportunity to "warm up" to
>cryostat temperature before attempting to section them. Leave your
>frozen blocks in the cryostat for 30-60 minutes before sectioning to
>allow them to come to optimum temperature
------------------------------
Message: 10
Date: Mon, 9 Aug 2010 09:00:29 -0700 (MST)
From: Adrienne Aperghis Kavanagh <aaperghis <@t> uspath.com>
Subject: [Histonet] Precipitate in Processor
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<12063462.318193.1281369629316.JavaMail.root <@t> mail3d.brinkster.com>
Content-Type: text/plain; charset=utf-8
Hello Everyone,
Has anyone ever seen a (salt?) precipitate in their alcohols following
formalin? While changing the processor this morning, I noticed a
precipitate in the 80% alcohol and 95% alcohol (NOT in the 70% alcohol).
It is white and grainy. The alcohols were otherwise unaffected.
We are using a 10% NBF containing:
Formaldehyde Water
Sodium Phosphate, monobasic
Sodium Phosphate, dibasic
Methanol
And our alcohols are all reagent grade.
Any help would be very much appreciated! Thank you in advance!
Adrienne Aperghis Kavanagh
US PATH
30 W. Century Road
Suite 255
Paramus NJ 07652
------------------------------
Message: 11
Date: Mon, 9 Aug 2010 12:13:18 -0400
From: Brandi Higgins <brandihiggins <@t> gmail.com>
Subject: Re: [Histonet] Precipitate in Processor
To: Adrienne Aperghis Kavanagh <aaperghis <@t> uspath.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<AANLkTi=dF7b97xeH7Y--R6Wh_J=ZBTBnoisYFvdFh29H <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Phosphate buffered formalin followed by concentrated alcohol will
produce these phosphate salts. To prevent this, formalin should be
followed by
alcohol of 70% or less. Also, when you change your processing solutions,
you can do a water flush (we do first 4 solutions - 2 formalin 2
alcohol) to
dissolve any salts that may be built up in the lines.
Brandi Higgins HT(ASCP), BS
On Mon, Aug 9, 2010 at 12:00 PM, Adrienne Aperghis Kavanagh <
aaperghis <@t> uspath.com> wrote:
> Hello Everyone,
>
> Has anyone ever seen a (salt?) precipitate in their alcohols following
> formalin? While changing the processor this morning, I noticed a
> precipitate in the 80% alcohol and 95% alcohol (NOT in the 70%
> alcohol). It
> is white and grainy. The alcohols were otherwise unaffected.
>
> We are using a 10% NBF containing:
> Formaldehyde Water
> Sodium Phosphate, monobasic
> Sodium Phosphate, dibasic
> Methanol
>
> And our alcohols are all reagent grade.
>
> Any help would be very much appreciated! Thank you in advance!
>
>
> Adrienne Aperghis Kavanagh
> US PATH
> 30 W. Century Road
> Suite 255
> Paramus NJ 07652
>
>
> _______________________________________________ Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 12
Date: Mon, 9 Aug 2010 12:18:53 -0400
From: "Pam Barker" <relia1 <@t> earthlink.net>
Subject: [Histonet] RELIA Solutions Hot Histology Job Alert!!
Histotech needed in Indianapolis area.
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <E1OiV3r-0006l1-TX <@t> elasmtp-mealy.atl.sa.earthlink.net>
Content-Type: text/plain; charset="iso-8859-1"
Hi Histonetters!
I want to put the word out on another position I am very excited about.
I am working with a hospital in the Indianapolis area who is in need of
a histotech with at least 5 years of experience for a permanent day
shift position. My client offers excellent pay, benefits and relocation
assistance. If you or anyone you know might be interested please
contact me at relia1 <@t> earthlink.net or toll free at 866-607-3542.
Thanks-Pam
Thank You!
Pam Barker
President RELIA
Specialists in Allied Healthcare Recruiting
5703 Red Bug Lake Road #330
Winter Springs, FL 32708-4969
Phone: (407)657-2027
Cell: (407)353-5070
FAX: (407)678-2788
E-mail: relia1 <@t> earthlink.net
www.facebook.com search Pam Barker RELIA
www.linkedin.com/reliasolutions www.myspace.com/pamatrelia
www.twitter.com/pamatrelia
------------------------------
Message: 13
Date: Mon, 9 Aug 2010 18:22:04 +0200
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] Re: [Special Stains] Masson's Trichrome
To: "'Sherwood, Margaret '" <MSHERWOOD <@t> PARTNERS.ORG>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <E1C77632E46B41F8AF2EC24E906E7967 <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
We don't perfom Masson Trichrome but the one-step Gomori version and
another rather similar trichrome called SFOG.
I use Bouins for one year, better said, when the reagens in the coplin
jar goes low, I fill it up again.
Our selfmade SFOG solution is used even longer than one year. And the
commercial Gomori-solution is used for a half year.
Weigert's is made once a week, but the results show that the older
solution doesn't work as well. Since nuclei aren't really important in
the trichromes, it doesn't matter that much.
It's important, that the pH is correct and low enough, to render the
solution stable.
Our numbers of stained trichromes are about 150 to 200 per year.
Gudrun
-----Urspr�ngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von
Sherwood, Margaret
Gesendet: Montag, 09. August 2010 16:33
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Re: [Special Stains] Masson's Trichrome
To all:
We keep having the same discussion re: special stains. How long can you
use them before discarding? I am specifically referring to the stains
used in
Masson's Trichrome.
We are a research lab and, therefore, don't run the volume that most
labs do.
We have found, for instance, that we can use the Weigert's Hematoxylin
(A&B) more than once, with filtering (even though it is stated it should
be made
fresh). We also use Bouin's Fix, Biebrich Scarlet and Aniline Blue more
than once. Is there a "set rule" when we should discard these solutions
(i.e. after
5X use or 1 month)?
I would appreciate some feedback from the experts!
Thanks!
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
The information in this e-mail is intended only for the person to whom
it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners
Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you
in error
but does not contain patient information, please contact the sender and
properly dispose of the e-mail.
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Message: 14
Date: Mon, 9 Aug 2010 09:23:22 -0700
From: "Jon St.Onge" <Jon.St.Onge <@t> dako.com>
Subject: RE: [Histonet] Precipitate in Processor
To: "Adrienne Aperghis Kavanagh" <aaperghis <@t> uspath.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<8B07D141BCDE434285DC12B3290E3FB304EE5635 <@t> exbackca.caus.dako.net>
Content-Type: text/plain; charset="UTF-8"
That is a phosphate precipitate most likely from your 10%NBF.
Try going into a 70% ETOH first followed by higher concentrations.
To see for yourself pour 10%NBF into graded alcohols- Precipitate is
easy to see with 100% ETOH
Jon Henry St. Onge
Dako North America
Quality Control Supervisor
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Adrienne
Aperghis Kavanagh
Sent: Monday, August 09, 2010 9:00 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Precipitate in Processor
Hello Everyone,
Has anyone ever seen a (salt?) precipitate in their alcohols following
formalin? While changing the processor this morning, I noticed a
precipitate in the 80% alcohol and 95% alcohol (NOT in the 70% alcohol).
It is white and grainy. The alcohols were otherwise unaffected.
We are using a 10% NBF containing:
Formaldehyde Water
Sodium Phosphate, monobasic
Sodium Phosphate, dibasic
Methanol
And our alcohols are all reagent grade.
Any help would be very much appreciated! Thank you in advance!
Adrienne Aperghis Kavanagh
US PATH
30 W. Century Road
Suite 255
Paramus NJ 07652
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Message: 15
Date: Mon, 9 Aug 2010 12:25:25 -0400
From: Drew Meyer <41dmb41 <@t> gmail.com>
Subject: Re: [Histonet] Precipitate in Processor
To: Adrienne Aperghis Kavanagh <aaperghis <@t> uspath.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<AANLkTimWiQh2tnaQpV_D2JU+cSTgCuqovnQWEA4z-mMJ <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
My guess is that either your 70% wasn't made up properly and was a
higher concentration or it's been so long since you've changed the
solution that
the water is fully saturated with the formalin salts. If it becomes a
regular problem, you might consider reducing your first alcohol's
concentration to 60% or even 50%. Good luck!
Drew
On Mon, Aug 9, 2010 at 12:00, Adrienne Aperghis Kavanagh <
aaperghis <@t> uspath.com> wrote:
> Hello Everyone,
>
> Has anyone ever seen a (salt?) precipitate in their alcohols following
> formalin? While changing the processor this morning, I noticed a
> precipitate in the 80% alcohol and 95% alcohol (NOT in the 70%
> alcohol). It
> is white and grainy. The alcohols were otherwise unaffected.
>
> We are using a 10% NBF containing:
> Formaldehyde Water
> Sodium Phosphate, monobasic
> Sodium Phosphate, dibasic
> Methanol
>
> And our alcohols are all reagent grade.
>
> Any help would be very much appreciated! Thank you in advance!
>
>
> Adrienne Aperghis Kavanagh
> US PATH
> 30 W. Century Road
> Suite 255
> Paramus NJ 07652
>
>
> _______________________________________________ Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 16
Date: Mon, 9 Aug 2010 12:32:41 -0400
From: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>
Subject: [Histonet] Re: Stability of Special Stains
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9703E24536 <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
I want to thank everyone who answered my inquiry into the stability of
special stains. And a special thanks to Rena Fail, for the link to the
University of
Rochester's Special Stains manual. It was very helpful. You have
answered my
questions.
Peggy
Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org
The information in this e-mail is intended only for the person to whom
it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners
Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you
in error
but does not contain patient information, please contact the sender and
properly dispose of the e-mail.
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