[Histonet] 1. LCM on decaled bone sections (Elizabeth M Heimrich
Pierre CHAUMAT
pierre.chaumat <@t> alphelys.com
Fri Jul 18 16:08:59 CDT 2008
Dear Elisabeth,
I think we have some starts of answers for you.
We are a french company that does promote and sell LCM system from Arcturus
since 99 and we have developped a strong expertise for that.
Also, as a matter of coincidence for the discussion, we have developped a
formalin free fixative marketed under the name of RCL2.
Multiple things I can say :
1- Formalin fixation will most probably annhilate your possibilities of
working with proper RNA and proteins far beyond decal
2- Working with frozen section does not leave you a good enought morphology
to work with when selecting specific cells during LCM
I would also encourage you to read some publications about RCL2 having
demonstrated its performance including preserving both morphology and
molecular content at a very high level.
RCL2 fixed specimens can be paraffin embedded and then stored at room temp.
J of Molecular Diagnostics. 2006 Vol.8 p.157-169 Delfour C - Boulle
N.pdf
If you need more details, please don't hesitate to contact me again whether
it is LCM protocols or of course RCL2 use...!
Best regards
Pierre
Pierre CHAUMAT
President & CEO
ALPHELYS SAS
Passage Paul Langevin
Ferme des Ebisoires
78370 PLAISIR, FRANCE
Tel +33 1 30 07 52 95
Fax +33 1 30 07 51 56
Cell +33 6 03 47 75 92
http://www.alphelys.com
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Today's Topics:
1. LCM on decaled bone sections (Elizabeth M Heimrich)
2. Re: borrelia burgdorferi protocol (Richard Cartun)
3. AW: [Histonet] borrelia burgdorferi protocol (Gudrun Lang)
4. AP Cerner Copath vs Sunquest Copath vs others (Cristian Britos)
5. Question on Aluminum Staining (Herrick, James L.)
6. Re: CD4 Fluorescent Staining (Gayle Callis)
7. Formlain containers (Richard Cartun)
8. Desmoglein 3 antibody (Patti Loykasek)
9. RE: Formlain containers (Martin, Gary)
10. RE: hcv and mastcells (Tony Henwood)
11. Re: ER,PR, HER2/Neu (Richard Cartun)
12. RE: ER,PR, HER2/Neu (McMahon, Loralee A)
13. IBF and prostates-your experience (Greg Dobbin)
14. RE: ER,PR, HER2/Neu (Jesus Ellin)
15. Re: IBF and prostates-your experience (Greg Dobbin)
16. Re: IBF and prostates-your experience (Rene J Buesa)
17. FW: [Histonet] Question on Aluminum Staining (Herrick, James L.)
----------------------------------------------------------------------
Message: 1
Date: Thu, 17 Jul 2008 14:01:16 -0400
From: Elizabeth M Heimrich <elizabeth.heimrich <@t> bms.com>
Subject: [Histonet] LCM on decaled bone sections
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <487F88EC.8010505 <@t> bms.com>
Content-Type: text/plain; charset="iso-8859-1"
Hello everyone,
I have a question regarding the doability of LCM on decaled rat joint
specimens and the subsequent RNA extraction and amplification. The rat
joints have not been harvested yet, so there is no fixative decided upon.
Is this a feasible procedure? Does the decalcification of the bone destroy
the RNA, or is the main issue the formaldehyde cross-linking the proteins if
using FFPE tissue. I have queried a few scientists around here and have
received conflicting answers, so I come to the net....hoping to resolve this
issue. Some say to dissect out the synovium and take cryo sections because
the Formaldehyde will interfere with the RNA signal. Others say that the
problem resides with the decal procedure itself. Any and all thoughts
welcome.
Thank you,
Beth
------------------------------
Message: 2
Date: Thu, 17 Jul 2008 15:00:19 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: Re: [Histonet] borrelia burgdorferi protocol
To: <gu.lang <@t> gmx.at>,<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <487F5E8302000077000042AD <@t> gwmail6.harthosp.org>
Content-Type: text/plain; charset=US-ASCII
I have been doing IHC for Borrelia burgdorferi for years and have never seen
a positive human case; only in animal tissue.
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 07/17/08 12:33 PM >>>
Hi all!
Can anyone provide me a borrelia burgdorferi protocol for ihc, preferable on
the Benchmark XT? We will purchase a polyclonal antibody from Acris. In the
datasheet is no hint about pretreatment or titer. So some information would
be helpful to start with.
Thanks in advance
Gudrun Lang
------------------------------
Message: 3
Date: Thu, 17 Jul 2008 21:09:26 +0200
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] borrelia burgdorferi protocol
To: "'Richard Cartun'" <Rcartun <@t> harthosp.org>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <37E7D6EF2B8D440DB16B613E1E6D7A87 <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
In spite of your "negative experiences" would you please share your protocol
with me?
And what do you think is the cause, why it doesn't work on human tissue?
Gudrun Lang
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
-----Urspr|ngliche Nachricht-----
Von: Richard Cartun [mailto:Rcartun <@t> harthosp.org]
Gesendet: Donnerstag, 17. Juli 2008 21:00
An: gu.lang <@t> gmx.at; histonet <@t> lists.utsouthwestern.edu
Betreff: Re: [Histonet] borrelia burgdorferi protocol
I have been doing IHC for Borrelia burgdorferi for years and have never seen
a positive human case; only in animal tissue.
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 07/17/08 12:33 PM >>>
Hi all!
Can anyone provide me a borrelia burgdorferi protocol for ihc, preferable on
the Benchmark XT? We will purchase a polyclonal antibody from Acris. In the
datasheet is no hint about pretreatment or titer. So some information would
be helpful to start with.
Thanks in advance
Gudrun Lang
------------------------------
Message: 4
Date: Thu, 17 Jul 2008 15:26:02 -0400
From: "Cristian Britos" <cbritos <@t> pathsrv.com>
Subject: [Histonet] AP Cerner Copath vs Sunquest Copath vs others
To: "Richard Cartun" <Rcartun <@t> harthosp.org>, <gu.lang <@t> gmx.at>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BF1A18A99963FA459E741057D0F47A8B40E21B <@t> psidc2.PSI.LOCAL>
Content-Type: text/plain; charset="us-ascii"
Would someone be willing to share their experiences about Sunquest copath as
their LIS system for pathology versus other systems?
Needless to say, we are looking for options.... (currently we have cerner
copath 3.0) Thanks, Cris
------------------------------
Message: 5
Date: Thu, 17 Jul 2008 15:16:09 -0500
From: "Herrick, James L." <Herrick.James <@t> mayo.edu>
Subject: [Histonet] Question on Aluminum Staining
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<D26523836B2D5B4290F7F82AEAC6EB4F117348 <@t> msgebe53.mfad.mfroot.org>
Content-Type: text/plain; charset="us-ascii"
Hi everyone,
We are having trouble running ASA (acid solochrome azurin) stains on human
iliac crest bone biopsies for the detection of aluminum deposits.
Our negatives are showing a lot of blue around the outer edges of the
specimen. Although we cannot see staining around the trabecular bone, we
should also not see it around the periphery of the bone specimen. The
positive, on the other hand, looks very good. Has anyone had experience with
this type of stain, and if so, would you happen to know what we may be doing
wrong?
The recipe we use for our stain is .2 gms. of Chromeazural B in 20mL of dH2O
with a pH @ 5.0 (We incubate at room temp for 18hrs.), destain with 95%
methanol, rinse in dH2O, dehydrate, clear and coverslip.
Also, another problem we have encountered in the past, is a gold discoloring
of the specimen. Have you seen this problem before, and if so, do you know
how to correct it?
Any help we can get with this dilemma is very much appreciated.
Thanks again,
Jim
------------------------------
Message: 6
Date: Thu, 17 Jul 2008 14:41:37 -0600
From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
Subject: Re: [Histonet] CD4 Fluorescent Staining
To: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <006801c8e84d$844c5520$6401a8c0 <@t> DHXTS541>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
Just to clarify exactly what you are doing - are you using a rat antiMouse
CD4 conjugated to Alexa 488? After I get the answer, I will elaborate. We
do successful CD4 immunofluorescent staining but never use a CD4 primary
direct, with the primary conjugated to the fluorphore. We have four
different ways of doing the staining.
Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT
----- Original Message -----
From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, July 17, 2008 9:38 AM
Subject: [Histonet] CD4 Fluorescent Staining
I am trying an antibody called Alexa Fluor 488 anti-mouse CD4 on fresh
frozen mouse tissue. Having only worked with AED/DAB staining, not very
familiar with this.
Does anyone have any experience with this and if you could send me some
tips/protocols. Thanks ahead of time
Loralee McMahon, HTL (ASCP)
ICC Supervisor
University of Rochester
Department of Pathology
(585) 275-7210
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------------------------------
Message: 7
Date: Thu, 17 Jul 2008 17:39:27 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: [Histonet] Formlain containers
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <487F83CF02000077000042C8 <@t> gwmail6.harthosp.org>
Content-Type: text/plain; charset=US-ASCII
Does anyone receive tissue specimens in plastic bags containing formalin?
We do and I don't like it. For some reason this client likes to use these
plastic bags instead of our standard formalin bottles. The bags have to be
cut open and are difficult to store. The person in the gross room ends up
placing all remaining tissue in one of our regular specimen bottles after
completing the gross examination. I am going to tell them that we are not
going to accept the specimens unless you can convince me that it is O.K.
(and I don't think that is going to happen). Thanks for your time.
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
------------------------------
Message: 8
Date: Thu, 17 Jul 2008 14:48:18 -0700
From: Patti Loykasek <ploykasek <@t> phenopath.com>
Subject: [Histonet] Desmoglein 3 antibody
To: histonet <histonet <@t> pathology.swmed.edu>
Message-ID: <C4A50C32.153E1%ploykasek <@t> phenopath.com>
Content-Type: text/plain; charset="US-ASCII"
Hi all. We have been trying to work up an antibody, desmoglein 3, in
squamous cell lung carcinoma. This is based on an article in Feb. 2007 Human
Pathology. We have had great difficulty in optimizing the antibody in FFPE.
If anyone else has this antibody up & working well, I would appreciate any
help with it. Thanks.
Patti Loykasek BS, HTL, QIHC
PhenoPath Laboratories
Seattle, WA
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------------------------------
Message: 9
Date: Thu, 17 Jul 2008 15:59:36 -0700
From: "Martin, Gary" <gmartin <@t> marshallmedical.org>
Subject: RE: [Histonet] Formlain containers
To: "Richard Cartun" <Rcartun <@t> harthosp.org>, "Histonet"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<6ED9D4252F278841A0593D3D788AF24C02DFF0D2 <@t> mailsvr.MARSHMED.local>
Content-Type: text/plain; charset="us-ascii"
We use both ... the bags have not been a problem ... as a matter for
fact I prefer them. They seem to store easier. We also noticed that
many times clients try to stuff a very large specimen into a small
bottle, and fixation can suffer. The bags we use are designed for
pathology use. They are manufactured by Bitran, and sold by Cardinal
Health
Gary
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Richard
Cartun
Sent: Thursday, July 17, 2008 2:39 PM
To: Histonet
Subject: [Histonet] Formlain containers
Does anyone receive tissue specimens in plastic bags containing
formalin? We do and I don't like it. For some reason this client likes
to use these plastic bags instead of our standard formalin bottles. The
bags have to be cut open and are difficult to store. The person in the
gross room ends up placing all remaining tissue in one of our regular
specimen bottles after completing the gross examination. I am going to
tell them that we are not going to accept the specimens unless you can
convince me that it is O.K. (and I don't think that is going to happen).
Thanks for your time.
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 10
Date: Fri, 18 Jul 2008 09:15:17 +1000
From: "Tony Henwood" <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] hcv and mastcells
To: <gu.lang <@t> gmx.at>, <histonet <@t> lists.utsouthwestern.edu>
Cc: TJJ <@t> Stowers-Institute.org
Message-ID: <B9EAF61856077F47BF9BE2F89AFC555202FAFF8E <@t> hedwig.nch.kids>
Content-Type: text/plain; charset="us-ascii"
Gudrun,
Teri Johnson [TJJ <@t> Stowers-Institute.org]gave an excellent summary of
this problem on the IHC resource groups listserver (Wednesday, 21 May
2008) and hopefully with her leave, I have reproduced it below:
"I did some rooting around in the literature and found out why you get
antibody staining (in negative control material) of mast cells, and what
you can do about it.
Summary During investigations of murine and human mast cell
immunoreactivity with potential anti-interleukin-4 antibodies,
non-specific, non-immunological labelling of mouse and human mast cells
became apparent. Non-specific, non-immunological labelling was
identified by (i) immunolabelling of mast cells when using control
isotype primary antibodies, (ii) ability of conjugated secondary
antibodies to label mast cells without prior mast cell exposure to a
primary antibody, (iii) extinction of the non-specific labelling and
retention of specific labelling when the pH of the diluting and washing
buffers is shifted from pH 7.2 to pH 6.0, and (iv) reduction/extinction
of the labelling when the antibodies are pre-incubated with soluble
heparin prior to immunostaining. The site of the reactivity on the
electron microscope level was shown to be confined to the mast cell
secretory granules. The results of this study support the hypothesis
that non-specific labelling of mast cells results from an ionic
interaction between the F(ab)2 segments of antibodies and the heparin
constituent of the mast cell secretory granules. This study points out
the necessity of stringent controls when using immunohistochemistry to
determine mast cell reactivity to various antibodies.
(Ref: Histochemical Journal, Vol. 25(9), Sept 1993, Schiltz et al)
An additional source for this (1989-1990 publication):
It is postulated that the granules of TMC bind certain antibodies by a
cation-exchange mechanism involving ionic interactions with positively
charged groups in the F(ab')2 and/or Fc segments. (J Histochem Cytochem
38:859-867, 1990)
And, finally, I found this little gem:
To avoid nonspecific binding of the antibodies to the heparin contained
in mast cells, the specificity of immunostaining was checked. PBS for
washing and dilution of the antibodies was acidified to pH 6.0.
Preadsorption of the STG I antibody with 1000 U/ml heparin (Sigma; St
Louis, MO) for 60 min at RT before use was carried out to avoid
nonspecific binding of immunoglobulins and heparin. The specificity of
the immunohistochemistry was confirmed with negative controls: absence
of primary or secondary antibody and avidin-labeled peroxidase. Normal
non-immune mouse serum was also used instead of the primary antibody.
(Ref: JHistochemCytochem 49(3): 341, Kumura et al)
Tammy: You never mentioned if you were getting mast cell staining in
your negative control. If no, then the above may not pertain to you and
you may have an issue of specific cross-reactivity with the antibody."
Good luck!
Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110
Thanks Terri,
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun
Lang
Sent: Friday, 18 July 2008 2:31 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] hcv and mastcells
Hi all!
I did an immunohistochemistry for heptatitis C virus on skin of a man,
who formerly had an hcv infection. The mastcells showed a granular
staining in the cytoplasm. Has anyone experience or explanations with
this phenomen? Could it be due to a cross-reaction or are there really
viruses (at least
particel) in the mastcell-granula?
Thanks in advance
Gudrun Lang
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Message: 11
Date: Fri, 18 Jul 2008 09:42:30 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: Re: [Histonet] ER,PR, HER2/Neu
To: <histonet <@t> lists.utsouthwestern.edu>, "Jesus Ellin"
<JEllin <@t> yumaregional.org>
Message-ID: <4880658602000077000042F7 <@t> gwmail6.harthosp.org>
Content-Type: text/plain; charset=US-ASCII
Why do you want to use image analysis?
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
>>> "Jesus Ellin" <JEllin <@t> yumaregional.org> 07/17/08 12:39 PM >>>
We are in the early stages of getting into ER/PR/HER2Neu scanning
technology using the Ventana Image Analysis System (VIAS). Wanting to
know the Pit falls and also what else to expect when going into this
realm,, Any information would greatly be appreciated.
Jesus Ellin HT/PA ASCP
Yuma Regional Medical Center
928-336-1743
This message is confidential, intended only for the named
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or exempt from disclosure under applicable law. If you are
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strictly prohibited. If you receive this message in error,
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Thank You.
_______________________________________________
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------------------------------
Message: 12
Date: Fri, 18 Jul 2008 09:50:05 -0400
From: "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
Subject: RE: [Histonet] ER,PR, HER2/Neu
To: "Richard Cartun" <Rcartun <@t> harthosp.org>,
<histonet <@t> lists.utsouthwestern.edu>, "Jesus Ellin"
<JEllin <@t> yumaregional.org>
Message-ID:
<2CF6F6B05263EA4EBAB07781B51E5DB0025A2810 <@t> e2k3ms1.urmc-sh.rochester.edu>
Content-Type: text/plain; charset="iso-8859-1"
We are in the process of purchasing a image analysis machine from DAKO
called the ACIS. I believe that the ACIS has a nicer interface with the
user, but I haven't had the opportunity to actually sit down and use the
Ventana machine myself. I have only seen it being used.
Loralee McMahon, HTL (ASCP)
ICC Supervisor
University of Rochester
Department of Pathology
(585) 275-7210
________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Richard Cartun
Sent: Fri 7/18/2008 9:42 AM
To: histonet <@t> lists.utsouthwestern.edu; Jesus Ellin
Subject: Re: [Histonet] ER,PR, HER2/Neu
Why do you want to use image analysis?
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
>>> "Jesus Ellin" <JEllin <@t> yumaregional.org> 07/17/08 12:39 PM >>>
We are in the early stages of getting into ER/PR/HER2Neu scanning
technology using the Ventana Image Analysis System (VIAS). Wanting to
know the Pit falls and also what else to expect when going into this
realm,, Any information would greatly be appreciated.
Jesus Ellin HT/PA ASCP
Yuma Regional Medical Center
928-336-1743
This message is confidential, intended only for the named
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or exempt from disclosure under applicable law. If you are
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Thank You.
_______________________________________________
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------------------------------
Message: 13
Date: Fri, 18 Jul 2008 11:24:47 -0300
From: "Greg Dobbin" <gvdobbin <@t> ihis.org>
Subject: [Histonet] IBF and prostates-your experience
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s8807d89.075 <@t> ihis.org>
Content-Type: text/plain; charset=US-ASCII
Hi folks,
If were to use IBF for the collection of prostate cores and they only
fixed in IBF for 3 hrs before being transferred to 10% formalin for
processing with everything else, are the benefits of better nuclear
morphology still likely to be realized?
Thank you.
Greg
Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385
"Success is not the key to happiness.
Happiness is the key to success.
If you love what you are doing,
you will be successful."
- Albert Schweitzer
-------------------------
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courriel ou encore de vous en servir, et vous devriez l'effacer
imm?diatement de votre syst?me informatique.
-------------------------
------------------------------
Message: 14
Date: Fri, 18 Jul 2008 07:51:14 -0700
From: "Jesus Ellin" <JEllin <@t> yumaregional.org>
Subject: RE: [Histonet] ER,PR, HER2/Neu
To: "Richard Cartun" <Rcartun <@t> harthosp.org>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<29BE166A2CF48D459853F8EC57CD37E8011D8960 <@t> EXCHANGECLUSTER.yumaregional.local
>
Content-Type: text/plain; charset="US-ASCII"
We will be doing in-house ER,PR, Her/Neu studies and report out results
for treatment, instead of sending out to reference labs for this
service, We are also looking out intergrating this platform with our
Pathology Information System, so that pathologist can use this tool
without having to go out of the current program,, early stages with this
development though.
Jesus Ellin HT/PA ASCP
Yuma Regional Medical Center
928-336-1743
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Message: 15
Date: Fri, 18 Jul 2008 11:51:24 -0300
From: "Greg Dobbin" <gvdobbin <@t> ihis.org>
Subject: Re: [Histonet] IBF and prostates-your experience
To: <Rcartun <@t> harthosp.org>,<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s88083d4.065 <@t> ihis.org>
Content-Type: text/plain; charset=US-ASCII
Hi Richard,
I always assume most people in Histology know more than I do, so I
figured IBF was well known in histo circles. Sorry. IBF is a commecial
fixative available from Surgipath, which contains isopropyl alcohol,
methanol, barium chloride, and low percent formalin (<3% by weight).
Cheers!
Greg
Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385
"Success is not the key to happiness.
Happiness is the key to success.
If you love what you are doing,
you will be successful."
- Albert Schweitzer
>>> "Richard Cartun" <Rcartun <@t> harthosp.org> 7/18/2008 11:39 AM >>>
Hi Greg:
What is "IBF"?
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
>>> "Greg Dobbin" <gvdobbin <@t> ihis.org> 07/18/08 10:24 AM >>>
Hi folks,
If were to use IBF for the collection of prostate cores and they only
fixed in IBF for 3 hrs before being transferred to 10% formalin for
processing with everything else, are the benefits of better nuclear
morphology still likely to be realized?
Thank you.
Greg
Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385
"Success is not the key to happiness.
Happiness is the key to success.
If you love what you are doing,
you will be successful."
- Albert Schweitzer
-------------------------
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Message: 16
Date: Fri, 18 Jul 2008 08:08:02 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] IBF and prostates-your experience
To: Histonet <@t> lists.utsouthwestern.edu, Greg Dobbin <gvdobbin <@t> ihis.org>
Message-ID: <580192.32789.qm <@t> web65708.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Probably, because in 3 hours in IBF the biopsies will probably be fixed with
the alcohols contained in IBF (methanol and 2-propanol). What I don't see is
why you want to transfer them to 10% NBF.
Reni J.
--- On Fri, 7/18/08, Greg Dobbin <gvdobbin <@t> ihis.org> wrote:
From: Greg Dobbin <gvdobbin <@t> ihis.org>
Subject: [Histonet] IBF and prostates-your experience
To: Histonet <@t> lists.utsouthwestern.edu
Date: Friday, July 18, 2008, 10:24 AM
Hi folks,
If were to use IBF for the collection of prostate cores and they only
fixed in IBF for 3 hrs before being transferred to 10% formalin for
processing with everything else, are the benefits of better nuclear
morphology still likely to be realized?
Thank you.
Greg
Greg Dobbin, R.T.
Chief Technologist, Histology Lab
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385
"Success is not the key to happiness.
Happiness is the key to success.
If you love what you are doing,
you will be successful."
- Albert Schweitzer
-------------------------
Statement of Confidentiality
This message (including attachments) may contain confidential or privileged
information intended for a specific individual or organization. If you have
received this communication in error, please notify the sender immediately.
If
you are not the intended recipient, you are not authorized to use, disclose,
distribute, copy, print or rely on this email, and should promptly delete
this
email from your entire computer system.
D?claration de confidentialit?
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confidentiels ? l'intention d'une personne ou d'un organisme
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veuillez
en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le
destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer,
distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et
vous
devriez l'effacer imm?diatement de votre syst?me informatique.
-------------------------
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Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 17
Date: Fri, 18 Jul 2008 11:50:18 -0500
From: "Herrick, James L." <Herrick.James <@t> mayo.edu>
Subject: FW: [Histonet] Question on Aluminum Staining
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<D26523836B2D5B4290F7F82AEAC6EB4F11734B <@t> msgebe53.mfad.mfroot.org>
Content-Type: text/plain; charset="us-ascii"
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Herrick,
James L.
Sent: Thursday, July 17, 2008 3:16 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Question on Aluminum Staining
Hi everyone,
We are having trouble running ASA (acid solochrome azurin) stains on
human iliac crest bone biopsies for the detection of aluminum deposits.
Our negatives are showing a lot of blue around the outer edges of the
specimen. Although we cannot see staining around the trabecular bone, we
should also not see it around the periphery of the bone specimen. The
positive, on the other hand, looks very good. Has anyone had experience
with this type of stain, and if so, would you happen to know what we may
be doing wrong?
The recipe we use for our stain is .2 gms. of Chromeazural B in 20mL of
dH2O with a pH @ 5.0 (We incubate at room temp for 18hrs.), destain with
95% methanol, rinse in dH2O, dehydrate, clear and coverslip.
Also, another problem we have encountered in the past, is a gold
discoloring of the specimen. Have you seen this problem before, and if
so, do you know how to correct it?
Any help we can get with this dilemma is very much appreciated.
Thanks again,
Jim
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End of Histonet Digest, Vol 56, Issue 21
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