[Histonet] IHC on paraformaldehyde-fixed
Tony Henwood
AnthonyH <@t> chw.edu.au
Sun Dec 7 16:05:41 CST 2008
Tf,
In answer to you email:
=20
No I do not carry toxic liquid in my car.
But does the PFA powder dissolve easily? My experience is that you need to =
make the solution alkaline then heat it.
I never add methanol to my 10% neutral buffered formalin (it is buffered an=
d diluted ie 10%). The risk of polymerisation of the formalin (since it is =
diluted) and formic acid formation (since it is buffered) is greatly reduce=
d.
=20
Regards=20
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20
Laboratory Manager & Senior Scientist=20
Tel: 612 9845 3306=20
Fax: 612 9845 3318=20
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20
-----Original Message-----
From: tf [mailto:tifei <@t> foxmail.com]=20
Sent: Saturday, 6 December 2008 3:01 PM
To: Reuel Cornelia; Tony Henwood; histonet; anh2006 <@t> med.cornell.edu; Jan S=
hivers
Subject: Re: RE: RE: [Histonet] IHC on paraformaldehyde-fixed
=09
=09
you want to carry a bottle of toxic liquid on your car? or you will take a=
box of powder that can dissolve into useful solution easily?
=20
You have to add methanol in 10% formalin & 4% formaldehyde, rather 4% para=
formaldhyde....PFA is methanol free..it's very important.
=20
=20
2008-12-06=20
=09
________________________________
tf=20
=09
________________________________
=B7=A2=BC=FE=C8=CB=A3=BA Reuel Cornelia=20
=B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-06 00:09:36=20
=CA=D5=BC=FE=C8=CB=A3=BA Tony Henwood; tifei <@t> foxmail.com; histonet; anh200=
6 <@t> med.cornell.edu; Jan Shivers=20
=B3=AD=CB=CD=A3=BA=20
=D6=F7=CC=E2=A3=BA RE: RE: [Histonet] IHC on paraformaldehyde-fixed=20
=09
=09
I have been curious about this discussion. we used 4% paraformaldehyde
for smaller biopsies only because it has a faster penetration to tissue
than 10% formalin. In all my IHC that I have done. I observe that doing
an IHC with 4% paraformaldehyde does not necessarily need antigen
retrieval in comparison to 10% formalin either it will be human or
animal tissue but this depends on how long was it fix, our 4%
paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6
to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can
comment on the effect on this to tissue if you say you will use 4%
paraformaldehyde for storage and transportation.=20
Reuel Cornelia, BS MT, AMT
Cellular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
Tel: 214-559-7766
fax: 214-559-7768
>>> "Tony Henwood" <AnthonyH <@t> chw.edu.au> 12/04/08 9:29 PM >>>
tf wrote:
=20
"I DO believe that one reason some people use 4% PFA rather 10%
formalin is that PFA is a bit more stable, both for storage and
transportation~~~."
=20
I have not heard this before.
Do you have a reference for this?
=20
=20
Regards=20
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20
Laboratory Manager & Senior Scientist=20
Tel: 612 9845 3306=20
Fax: 612 9845 3318=20
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20
-----Original Message-----
From: tf [mailto:tifei <@t> foxmail.com]=20
Sent: Friday, 5 December 2008 2:11 PM
To: Tony Henwood; anh2006 <@t> med.cornell.edu; Jan Shivers;
histonet
Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed
the basic principles are the same for most cross-linking
fixatives and induce similar bonds=20
the difference you observed between may due to any other
variability, or the co-fixative you used.
=20
I DO believe that one reason some people use 4% PFA rather 10%
formalin is that PFA is a bit more stable, both for storage and
transportation~~~.
=20
=20
=20
=20
2008-12-05=20
________________________________
tf=20
________________________________
=B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood=20
=B7=A2=CB=CD=BC=E4=A3=BA 2008-12-05 06:00:03=20
=CA=D5=BC=FE=C8=CB=A3=BA anh2006 <@t> med.cornell.edu; Jan Shivers; histonet=20
=B3=AD=CB=CD=A3=BA=20
=D6=F7=A3=BA RE: [Histonet] IHC on paraformaldehyde-fixed=20
Interesting point.
Since 10% buffered formalin (made from the concentrated 38%
formaldehyde) contain about 1% methanol, has it been shown that
this has
a deleterious effect on ANY antigens or are we expecting this
worse case
senario as being the norm?
I am not aware of any antigens (or antigen-antibody combination)
that
has been badly effected by 10% formalin that is NOT effected by
10%
formaldehyde. Are you aware of any??
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead=20
Cnr Hawkesbury Road and Hainsworth Street, Westmead=20
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20
-----Original Message-----
From: anh2006 <@t> med.cornell.edu [mailto:anh2006 <@t> med.cornell.edu]=20
Sent: Friday, 5 December 2008 1:31 AM
To: Tony Henwood; Jan Shivers; histonet
Subject: Re: [Histonet] IHC on paraformaldehyde-fixed
So true. However, be aware that 10% neutral buffered formalin we
use has
methanol in it which may affect certain antigens so there may be
some
difference in staining (hence why for mouse work we now only use
4% PFA
in pure PBS). It is good to be aware of the other ingredients in
your
fixative solutions, whether commercially prepared or a homemaede
recipe,
as it isn't only the formaldehyde fixative which can make a
difference.
-----Original Message-----
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Date: Thu, 04 Dec 2008 09:35:09=20
To: Jan Shivers<shive003 <@t> umn.edu>;
histonet<histonet <@t> lists.utsouthwestern.edu>
Subject: RE: [Histonet] IHC on paraformaldehyde-fixed
Gee I hate the term paraformaldehyde (as many of you probably
know)
This is an example of how confusion of terms can cause
unnecessary work.
Is "4% paraformaldehyde" different from 4 % formaldehyde?
No
Should any procedure done to tissues fixed in "4%
paraformaldehyde" give
results different to those fixed in 4% formaldehyde or 10%
formalin?=20
No since they are the same thing.
As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002)
state
when paraformaldehyde actually becomes a fixative, it is no
longer
paraformaldehyde by chemistry or fixation capacity. Rather, it
is
formaldehyde in water without methanol or any other stabiliser.
Without
heat and an alkaline environment, paraformaldehyde in water is
simply a
paraformaldehyde suspension with little fixation capacity. If
the
fixative is prepared from paraformaldehyde then it should be
termed 4%
formaldehyde freshly prepared from paraformaldehyde. If a
concentrated
formalin solution (40% formaldehyde) is used, then it should be
termed
10% formalin.
If you do a search on Histonet for paraformaldehye, you will
find that
this topic has been extensively discussed.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory
Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead=20
Cnr Hawkesbury Road and Hainsworth Street, Westmead=20
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu=20
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Jan
Shivers
Sent: Thursday, 4 December 2008 8:34 AM
To: histonet
Subject: [Histonet] IHC on paraformaldehyde-fixed
Has anyone ever done IHC on parafomaldehyde-fixed tissues, and
if so,
how well did it work? Will the same antigen-retrieval methods
used with
formalin-fixed tissue be applicable?
I'm asking for an investigator, who already has his tissues
fixed in
paraformaldehyde.
Jan Shivers
Senior Scientist
Pathology Teaching Program
Histology/IHC/EM Section Head
University of Minnesota
Veterinary Diagnostic Laboratory
1333 Gortner Ave.
St. Paul, MN 55108
612-624-7297
shive003 <@t> umn.edu_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu=20
http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20
*********************************************************************
This email and any files transmitted with it are confidential
and
intended solely for the use of the individual or entity to whom
they are
addressed. If you are not the intended recipient, please delete
it and
notify the sender.
Views expressed in this message and any attachments are those of
the
individual sender, and are not necessarily the views of The
Children's
Hospital at Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected,
The
Childrens Hospital at Westmead accepts no liability for any
consequential damage resulting from email containing computer
viruses.
**********************************************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu=20
http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20
*********************************************************************
This email and any files transmitted with it are confidential
and intended solely for the use of the individual or entity to whom they
are addressed. If you are not the intended recipient, please delete it
and notify the sender.
Views expressed in this message and any attachments are those of
the individual sender, and are not necessarily the views of The
Children's Hospital at Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected,
The Childrens Hospital at Westmead accepts no liability for any
consequential damage resulting from email containing computer viruses.
**********************************************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu=20
*********************************************************************
This email and any files transmitted with it are confidential and
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient, please delete it and
notify the sender.
Views expressed in this message and any attachments are those of the
individual sender, and are not necessarily the views of The Children's
Hospital at Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The
Childrens Hospital at Westmead accepts no liability for any
consequential damage resulting from email containing computer viruses.
**********************************************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu=20
http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20
**************************************************************************=
*****************************************=20
Texas Scottish Rite Hospital for Children is one of the nation's leading p=
ediatric centers for the=20
treatment of orthopedic conditions, certain related neurological disorders=
and learning disorders,=20
such as dyslexia. This email transmission and/or its attachments may cont=
ain confidential health=20
information, intended only for the use of the individual or entity named a=
bove.=20
The authorized recipient of this information is prohibited from disclosing=
it to any other party unless=20
required to do so by law and is required to delete/destroy the information=
after its stated need has
been fulfilled. If you are not the intended recipient, any disclosure, co=
pying, distribution or action=20
taken in reliance on the contents of this email transmission is prohibited=
. If you have received this
information in error, please notify the sender immediately and delete this=
information.=20
We appreciate your efforts to protect the children's confidential informat=
ion.=20
**************************************************************************=
*****************************************=20
*********************************************************************
This email and any files transmitted with it are confidential and intended =
solely for the use of the individual or entity to whom they are addressed. =
If you are not the intended recipient, please delete it and notify the send=
er.
Views expressed in this message and any attachments are those of the indivi=
dual sender, and are not necessarily the views of The Children's Hospital a=
t Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens=
Hospital at Westmead accepts no liability for any consequential damage res=
ulting from email containing computer viruses.
**********************************************************************
More information about the Histonet
mailing list