[Histonet] RE: NSH Teleconference Today
Carrie Diamond
Carrie <@t> nsh.org
Wed May 28 10:43:07 CDT 2008
Hello -
If you have questions regarding the NSH Teleconference, please contact the NSH office at 443.535.4060 or via email at histo <@t> nsh.org.
Thank you,
Carrie Diamond
Executive Director
National Society for Histotechnology
10320 Little Patuxent Parkway
Suite 804
Columbia, MD 21044
P: 443.535.4060
Direct: 443.535.4066
Fax: 443.535.4055
E-mail: carrie <@t> nsh.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Wednesday, May 28, 2008 11:34 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 54, Issue 41
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Today's Topics:
1. Re: IF question (Richard Cartun)
2. RE: Biotin free detection of goat primary antibodies
(C.M. van der Loos)
3. Thick section penetration for IHC (Bob Nienhuis)
4. Shirley Phua is out-of-office ... (Shirley PHUA)
5. RE: Biotin free detection of goat primary antibodies (Sally Price)
6. evaluation of new antibody cap question 22750
(Warren_Eddings <@t> ssmhc.com)
7. KOH for nails (dcojita <@t> tampabay.rr.com)
8. Rate of formalin penetration in human brain sections
(Karen_Skish <@t> rush.edu)
9. RE: IF question (Mickie Johnson)
10. Reaction between Haemoglobin and acetic acid (Tony Henwood)
11. RE: Reaction between Haemoglobin and acetic acid (kemlo)
12. nerve bx (Heath, Nancy L.)
13. Re: Rate of formalin penetration in human brain sections
(Rene J Buesa)
14. Re: KOH for nails (Rene J Buesa)
15. Re: Reaction between Haemoglobin and acetic acid (Rene J Buesa)
16. Cryostat in the Washington, DC area? (Norman Meres)
17. heat + formalin (Nancy Schmitt)
18. Re: Rate of formalin penetration in human brain sections
(Geoff McAuliffe)
19. Re: Rate of formalin penetration in human brain sections
(Bryan Hewlett)
20. NSH teleconference today (Ingles Claire)
21. RE: heat + formalin (Juan Gutierrez)
22. Re: heat + formalin (Rene J Buesa)
23. Formalin alternative fixative for rodent and large animal
(stephanie.d.rivera <@t> gsk.com)
24. Experiencing Chatter in paraffin ribbons (Nicole C Walsh)
25. RE: NSH teleconference today (Mahoney,Janice A)
26. RE: Rate of formalin penetration in human brain sections
(Marshall Terry Dr, Consultant Histopathologist)
----------------------------------------------------------------------
Message: 1
Date: Tue, 27 May 2008 13:26:41 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: Re: [Histonet] IF question
To: "Anne van Binsbergen" <annigyg <@t> gmail.com>,
"histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>,
<histonet-request <@t> lists.utsouthwestern.edu>
Message-ID: <483C0C110200007700002CF7 <@t> gwmail6.harthosp.org>
Content-Type: text/plain; charset=US-ASCII
We use tonsil, freshly cut.
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
>>> "Anne van Binsbergen" <annigyg <@t> gmail.com> 05/27/08 12:11 AM >>>
Hi Histonetters
is anyone out there running IF controls - specifically for renals, skins
if so what and how?
please advise asap
thanks
Anne
--
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------------------------------
Message: 2
Date: Tue, 27 May 2008 19:56:28 +0200
From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
Subject: [Histonet] RE: Biotin free detection of goat primary
antibodies
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <e1dd87176dc91f84.483c676c <@t> amc.uva.nl>
Content-Type: text/plain; charset=us-ascii
Dear Min-Han,We solved this problem with a 3-step approach: goat primary - rabbit anti-goat IgG - anti-rabbit polymer/HRP. The second step we purchased from Southern Biotech Associates, Birmingham, AL. Dilution of the rabbit anti-goat IgG (human IgG adsorbed) came out at 1:2000-5000 (30 min, RT). Any anti-rabbit polymer/HRP will do. Gayle is fully right with respect to the negative control: find a non-immune goat IgG with a known IgG protein concentration. You need that to calculate the final dilution.Hope this helps a bit.Cheers, ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The NetherlandsDate: Tue, 27 May 2008 10:55:59 +0800
From: "Min-Han Tan" <minhan.tan <@t> gmail.com>
Subject: [Histonet] Biotin free detection of goat primary antibodies
To: histonet <@t> lists.utsouthwestern.edu
Dear all,
I would like to enquire - I am using a goat primary antibody currently, and
my negative control (omitting primary antibody) is showing background
staining - this is likely a result of biotin.
I'd like to enquire if any one is familiar with biotin free systems that can
detect goat antibodies. A search of the Dako / Envision product line does
not yield any results. I am very reluctant to add an avidin-biotin blocking
step to what is already an overnight incubation.
Thanks!
--
/min-han
------------------------------
Message: 3
Date: Tue, 27 May 2008 10:04:52 -0800
From: "Bob Nienhuis" <bob.nienhuis <@t> gmail.com>
Subject: [Histonet] Thick section penetration for IHC
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<45109da50805271104j20ca621ar7abcbf0ddf4f38fb <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
It has been suggested that the ABC detection system may not be the optimal
choice for IHC in thick brain sections (~100 microns). Apparently, the large
avidin-biotin complexes may not penetrate tissue too well.
What would you suggest instead, LSAB, polymer or...?
Bob Nienhuis
Neurobiology Research
UCLA / VA Medical Center
Los Angeles
------------------------------
Message: 4
Date: Wed, 28 May 2008 02:03:44 +0800
From: Shirley PHUA <Shirley_PHUA <@t> hsa.gov.sg>
Subject: [Histonet] Shirley Phua is out-of-office ...
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<OFDE20C9A4.00046D02-ON48257456.0063383E-48257456.0063383E <@t> gems2.gov.sg>
Content-Type: text/plain; charset=US-ASCII
I will be out of the office from 27-05-2008 to 28-05-2008.
I'll be away on 27 May 2008 afternoon.
Pathologists:
I will process your requests when I return. If urgent, please forward your
email to Henry_Kyaw <@t> hsa.gov.sg
------------------------------
Message: 5
Date: Tue, 27 May 2008 16:34:55 -0400
From: "Sally Price" <sprice2003 <@t> gmail.com>
Subject: RE: [Histonet] Biotin free detection of goat primary
antibodies
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<ffd26fa80805271334r3c1f00c5s7abd01f7648558f3 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Min-Han:
I think you're best bet is to try Biocare Medical's anti-Goat polymer
detection system -- which can be seen at:
http://www.biocaremed.com/promark.html. It is very easy to use and yeilds
very clean stains.
Cheers,
Sally
------------------------------
Message: 4
Date: Tue, 27 May 2008 10:55:59 +0800
From: "Min-Han Tan" <minhan.tan <@t> gmail.com>
Subject: [Histonet] Biotin free detection of goat primary antibodies
To: histonet <@t> lists.utsouthwestern.edu
Dear all,
I would like to enquire - I am using a goat primary antibody currently, and
my negative control (omitting primary antibody) is showing background
staining - this is likely a result of biotin.
I'd like to enquire if any one is familiar with biotin free systems that can
detect goat antibodies. A search of the Dako / Envision product line does
not yield any results. I am very reluctant to add an avidin-biotin blocking
step to what is already an overnight incubation.
Thanks!
------------------------------
Message: 6
Date: Tue, 27 May 2008 15:59:04 -0500
From: Warren_Eddings <@t> ssmhc.com
Subject: [Histonet] evaluation of new antibody cap question 22750
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OFC4C62E34.6CF11EDA-ON86257456.0073455C-86257456.00734568 <@t> SSMHC>
Content-Type: text/plain; charset="ISO-8859-1"
if useing prediluted antibodys
thanks
warren_eddings <@t> ssmhc.co _________________________________________________________________
Confiden attachments, is for the s may contain confidential and privi unauthorized review, use, disclosure or distributi prohibited. If you are not the intended recipient, please contact t he sender by reply email and destroy all copies of the original
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------------------------------
Message: 7
Date: Tue, 27 May 2008 17:11:31 -0400
From: <dcojita <@t> tampabay.rr.com>
Subject: [Histonet] KOH for nails
To: "'histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>,
<histonet-request <@t> lists.utsouthwestern.edu>
Message-ID:
<!~!UENERkVCMDkAAQACAAAAAAAAAAAAAAAAABgAAAAAAAAAdq7zShuC4ki+F530qNQCZMKAAAAQAAAAEq+/icCGiUePkQSImEp+7wEAAAAA <@t> tampabay.rr.com>
Content-Type: text/plain; charset="us-ascii"
Does anyone have a procedure they would be willing to share for KOH (looking
for nail fungus)?
Is this considered an AP or a clinical test?
What is the CPT code?
Any information you have would be greatly appreciated!
------------------------------
Message: 8
Date: Tue, 27 May 2008 15:45:24 -0500
From: Karen_Skish <@t> rush.edu
Subject: [Histonet] Rate of formalin penetration in human brain
sections
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF5D3E28E1.AF7FB3EE-ON86257456.00716652-86257456.00720614 <@t> rsh.net>
Content-Type: text/plain; charset="US-ASCII"
Hi--
One of our investigators is interested in the approximate rate of fixation
of human brain tissue, independent of any formaldehyde diffusion effects.
In other words, in a very small or very thin piece of human brain tissue,
what is the fixation rate? He found published data for rat kidney, but
would like to try to at least determine if the fixation rate should be
higher or lower in human brain tissue. He is looking for data for room
temperature, but any information would be greatly appreciated.
Thanks!
Karen M Skish, MS, PA(ASCP)MT
Pathologists' Assistant & Manager, Neuropathology Lab
Rush Alzheimer's Disease Center
Cohn Research Building, Lab 441
1735 West Harrison Street
Chicago IL 60612
------------------------------
Message: 9
Date: Tue, 27 May 2008 18:22:56 -0700
From: "Mickie Johnson" <mickie25 <@t> netzero.net>
Subject: RE: [Histonet] IF question
To: "'histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<!&!AAAAAAAAAAAYAAAAAAAAAGSBPkiRJN1Ck0zqFBQmG77CgAAAEAAAALBRd5cS9y1Ngco2Wriki6QBAAAAAA==@netzero.net>
Content-Type: text/plain; charset="us-ascii"
Hello Histonetters,
I saw that Jan Mahoney gave a talk on Lean Histology a couple of weeks ago
at the Illinois Histology Society seminar and was wondering if she would
give me a shout as I have someone who is interested in what the buzz is
about. Me too for that matter. Thanks Jan!
Best Regards,
Mickie
Mickie Johnson, B.S., HTL(ASCP)
Mohs Histology Consulting Services, LLC
& Mohs Lab Staffing
2507 S. Manito Blvd.
Spokane, WA 99203
509-954-7134
FAX 509-624-3926
Web: www.mohshistogyconsulting.com & www.mohslabstaffing.com
Email: mickie25 <@t> netzero.net
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------------------------------
Message: 10
Date: Wed, 28 May 2008 14:10:12 +1000
From: "Tony Henwood" <AnthonyH <@t> chw.edu.au>
Subject: [Histonet] Reaction between Haemoglobin and acetic acid
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <B9EAF61856077F47BF9BE2F89AFC555202FAFF02 <@t> hedwig.nch.kids>
Content-Type: text/plain; charset="us-ascii"
Hi all,
I have been asked what is the altered haemoglobin called that results
from the treatment of tissues with acetic acid.
All I could think of was that carbon monoxide reacts with haemoglobin to
form carboxyhemoglobin.
Any ideas?
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
*********************************************************************
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Message: 11
Date: Wed, 28 May 2008 09:11:42 +0100
From: "kemlo" <kemlo <@t> f2s.com>
Subject: RE: [Histonet] Reaction between Haemoglobin and acetic acid
To: "'Tony Henwood'" <AnthonyH <@t> chw.edu.au>,
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <72927951B67343EE84B30090C88C3D2B <@t> KemloPC>
Content-Type: text/plain; charset="US-ASCII"
SUMMARY
It is shown by sedimentation velocity measurements that the
presence of undissociated acetic acid favors the dissociation of
hemoglobin in a pH region in which hydrogen ion had previously
been thought to have the dominant effect.
Acknowledgments-We wish to acknowledge continuous discussion
and collaboration with Professors J. Wyman and E. Antonini
during the course of this work.
REFEREYCES
1. REITHEL, F. J., Advances in Protein Chem., 18, 123 (1963).
2. PEDERSEN, K. O., AND ANDERSSON, K. J. I., in T. SVEDBERG
AND K. 0. PEDERSEN (Editors), The ultracentrifuge, Oxford
University Press, London, 1940, p. 407.
3. ROSSI-FANELLI, A., ANTONINI, E., AND CAPUTO, A., Advances
in Protein Chem., 19, 73 (1964).
4. PERUTZ, M. F., Biochem. J., 94, 21P (1965).
5. CULLIS, A. F., MUIRHEAD, H., PERUTZ, M. F., ROSSMANN,
M. G., AND NORTH, A. C. T., Proc. Roy. Sot. London, Ser. A,
265, 161 (1962).
6. PHELPS, R. A., AND CANN, J. R., J. Am. Chem. Sot., 78, 3539
(1956).
7. CANN, J. R., J. Biol. Chem., 235, 2810 (1960).
8. CANN, J. R., AND GOAD, W. B., J. Biol. Chem., 240, 148 (1965).
9. FIELD, E. O., AND O'BRIEN, J. R., Bioehem. J., 60, 656 (1955).
10. ROSSI-FANELLI, A., ANTONINI, E., AND CAPUTO, A., J. Biol.
Chem., 236, 391 (1961).
11. LONG, C. (Editor), Biochemists' handbook, Spon Ltd., London,
1961.
12. PEDERSEN, K. O., J. Phys. Chem., 62, 1282 (1958).
Downloaded from www.jbc.org by on May 28, 2008
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tony Henwood
Sent: 28 May 2008 05:10
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Reaction between Haemoglobin and acetic acid
Hi all,
I have been asked what is the altered haemoglobin called that results
from the treatment of tissues with acetic acid.
All I could think of was that carbon monoxide reacts with haemoglobin to
form carboxyhemoglobin.
Any ideas?
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
*********************************************************************
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If you are not the intended recipient, please delete it and notify the
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Views expressed in this message and any attachments are those of the
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Hospital at Westmead
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------------------------------
Message: 12
Date: Wed, 28 May 2008 06:52:00 -0400
From: "Heath, Nancy L." <NHeath <@t> Lifespan.org>
Subject: [Histonet] nerve bx
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<130E8991F210424096EFC6F42EA33B2402B63361 <@t> LSCOEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain; charset="us-ascii"
Hi,
Does anyone know of a procedure for nerve biopsies/nerve teasing that
does not use Osmium Tetroxide??
Tx,
Nancy
------------------------------
Message: 13
Date: Wed, 28 May 2008 05:28:37 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Rate of formalin penetration in human brain
sections
To: Karen_Skish <@t> rush.edu, histonet <@t> lists.utsouthwestern.edu
Message-ID: <216952.98289.qm <@t> web65705.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
I think that this is a very interesting subject for your investigator to research by himself/herself (especially independent of any formaldehyde diffusion effects)!
René J.
Karen_Skish <@t> rush.edu wrote: Hi--
One of our investigators is interested in the approximate rate of fixation
of human brain tissue, independent of any formaldehyde diffusion effects.
In other words, in a very small or very thin piece of human brain tissue,
what is the fixation rate? He found published data for rat kidney, but
would like to try to at least determine if the fixation rate should be
higher or lower in human brain tissue. He is looking for data for room
temperature, but any information would be greatly appreciated.
Thanks!
Karen M Skish, MS, PA(ASCP)MT
Pathologists' Assistant & Manager, Neuropathology Lab
Rush Alzheimer's Disease Center
Cohn Research Building, Lab 441
1735 West Harrison Street
Chicago IL 60612
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 14
Date: Wed, 28 May 2008 05:33:23 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] KOH for nails
To: dcojita <@t> tampabay.rr.com, "'histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>,
histonet-request <@t> lists.utsouthwestern.edu
Message-ID: <895728.9462.qm <@t> web65706.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
KOH is used (at 10% for 30 minutes) to soften nails so they can be properly infiltrated and cut. That is an AP procedure (although it is NOT a decalcification code I always used CPT code 88331).
Nail fungus will require a fungus stain (I always used PAS) and this is an AP test for organisms identification (CPT code 88312)
René J.
dcojita <@t> tampabay.rr.com wrote:
Does anyone have a procedure they would be willing to share for KOH (looking
for nail fungus)?
Is this considered an AP or a clinical test?
What is the CPT code?
Any information you have would be greatly appreciated!
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 15
Date: Wed, 28 May 2008 05:40:44 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Reaction between Haemoglobin and acetic acid
To: Tony Henwood <AnthonyH <@t> chw.edu.au>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID: <113899.56959.qm <@t> web65716.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
In dilute solution (1%) acetic acid destroys red blood cells to facilitate the examination of white blood cells. At this conc. it separates the dermis from the epidermis.
René J.
Tony Henwood <AnthonyH <@t> chw.edu.au> wrote:
Hi all,
I have been asked what is the altered haemoglobin called that results
from the treatment of tissues with acetic acid.
All I could think of was that carbon monoxide reacts with haemoglobin to
form carboxyhemoglobin.
Any ideas?
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
*********************************************************************
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.
**********************************************************************
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Histonet <@t> lists.utsouthwestern.edu
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------------------------------
Message: 16
Date: Wed, 28 May 2008 09:52:55 -0400
From: Norman Meres <nmeres <@t> gmu.edu>
Subject: [Histonet] Cryostat in the Washington, DC area?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <F54EEE07-CCFD-44C9-ACC2-59A7E0B44204 <@t> gmu.edu>
Content-Type: text/plain; charset=US-ASCII; format=flowed; delsp=yes
Hello:
The cryostat that I was planning on using for my research is not
available at the moment. Is there anyone in the Washington, DC area
that would have one that I could use? I am sectioning epidermis of
lobsters.
Thanks,
Norman Meres
Doctoral Candidate
Environmental Science and Public Policy
George Mason University
------------------------------
Message: 17
Date: Wed, 28 May 2008 09:08:47 -0500
From: Nancy Schmitt <nancy_schmitt <@t> pa-ucl.com>
Subject: [Histonet] heat + formalin
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<9FC023A4AB52BB4D87DC6456081A822C087CA1 <@t> mercury.pa-ucl.com>
Content-Type: text/plain; charset="iso-8859-1"
Good Morning
Need some input, please. Does anyone combine heat during the formalin part
of processing? We are having some issues with breast cases that need to be
read out the next am - we are running them on a shorter processing schedule
(done at 4 am) - and not having the best results.We use a closed system
Tissue-Tek VIP 3000. Is anybody using microwave fixation?
I think we will just put them on the longer processor (done at 0630) and be
done - but wanted to check for other options.
Thanks in advance
Nancy Schmitt
Histology Coordinator
Dubuque, IA
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Message: 18
Date: Wed, 28 May 2008 10:19:20 -0400
From: Geoff McAuliffe <mcauliff <@t> umdnj.edu>
Subject: Re: [Histonet] Rate of formalin penetration in human brain
sections
To: Karen_Skish <@t> rush.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <483D69E8.8020506 <@t> umdnj.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Hi Karen:
Formalin fixes tissue slowly, even very thin slices. This has been
known for many, many years. The work of Medwar and of John R. Baker
(Principles of Biological Technique, John Wiley and Sons, 1958) comes to
mind. Perhaps there are slight differences between kidney and brain but
my guess is that if there is a difference it is insignificant. Before
trying to design (how are you going to define fixation?) and perform
such investigations I suggest a trip to the library. I doubt if the
information you seek is on line, it is too old. However, do not confuse
"old" with "out dated" or "bad". Good luck.
Geoff
Karen_Skish <@t> rush.edu wrote:
> Hi--
> One of our investigators is interested in the approximate rate of fixation
> of human brain tissue, independent of any formaldehyde diffusion effects.
> In other words, in a very small or very thin piece of human brain tissue,
> what is the fixation rate? He found published data for rat kidney, but
> would like to try to at least determine if the fixation rate should be
> higher or lower in human brain tissue. He is looking for data for room
> temperature, but any information would be greatly appreciated.
> Thanks!
> Karen M Skish, MS, PA(ASCP)MT
> Pathologists' Assistant & Manager, Neuropathology Lab
> Rush Alzheimer's Disease Center
> Cohn Research Building, Lab 441
> 1735 West Harrison Street
> Chicago IL 60612
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
**********************************************
------------------------------
Message: 19
Date: Wed, 28 May 2008 10:29:38 -0400
From: "Bryan Hewlett" <bhewlett <@t> cogeco.ca>
Subject: Re: [Histonet] Rate of formalin penetration in human brain
sections
To: <histonet <@t> lists.utsouthwestern.edu>, <Karen_Skish <@t> rush.edu>
Message-ID: <001301c8c0cf$430e19e0$6700a8c0 <@t> mainbox>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
Karen,
The formaldehyde fixation rate of tissue is determined from a combination of
the penetration rate, which is governed by diffusibilty,
the maximal covalent formaldehyde binding time, which is governed by the
formaldehyde 'clock' reaction, and the slow subsequent cross-linking which
then occurs,
This slow cross-linking is thought to be almost complete by 7 days post
formaldehyde binding time, but can continue over longer time periods.
What your investigator has found, is probably the published data related to
the maximal covalent formaldehyde binding time, i.e. 24 hours at room
temperature
as determined by Fox et al (reference #1). This study was essentially
independent of diffusibilty, i.e. negligible penetration time, since 16
micrometer thick sections of rat kidney were the substrate.
However, this study does not take into account subsequent further
cross-linking.
A study by Helander (reference #2) determined the maximal covalent
formaldehyde binding time as 25 hours.
This was performed on 4 mm thick slices of rabbit liver, so diffusibility
has to be taken into account.
In addition, this study also partially looked at subsequent further
cross-linking in relationship to reversibility.
A later study by Helander (reference #3) compared the maximal covalent
formaldehyde binding time between kidney and brain tissues.
This was stated to be 50 hours however, the tissues thickness was increased
to 8 mm and so the diffusibilty effect has to be taken into account even
more.
Reference #1.
Fox CH., et.al. Formaldehyde fixation. J Histochem. Cytochem. 1985; 33,
845 -853
Reference #2
Helander, KG. Kinetic studies of formaldehyde binding in tissue.
Biotechnique and Histochemistry. 1994; 69, 177 -179
Reference #3
Helander, K.G. Formaldehyde binding in Brain and Kidney: A kinetic study of
fixation.
The Journal of Histotechnology. 1999; 22(4), 317-318.
Regards,
Bryan
----- Original Message -----
From: <Karen_Skish <@t> rush.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, May 27, 2008 4:45 PM
Subject: [Histonet] Rate of formalin penetration in human brain sections
> Hi--
> One of our investigators is interested in the approximate rate of fixation
> of human brain tissue, independent of any formaldehyde diffusion effects.
> In other words, in a very small or very thin piece of human brain tissue,
> what is the fixation rate? He found published data for rat kidney, but
> would like to try to at least determine if the fixation rate should be
> higher or lower in human brain tissue. He is looking for data for room
> temperature, but any information would be greatly appreciated.
> Thanks!
> Karen M Skish, MS, PA(ASCP)MT
> Pathologists' Assistant & Manager, Neuropathology Lab
> Rush Alzheimer's Disease Center
> Cohn Research Building, Lab 441
> 1735 West Harrison Street
> Chicago IL 60612
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 20
Date: Wed, 28 May 2008 09:30:08 -0500
From: "Ingles Claire" <CIngles <@t> uwhealth.org>
Subject: [Histonet] NSH teleconference today
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<08A0A863637F1349BBFD83A96B27A50A120138 <@t> uwhis-xchng3.uwhis.hosp.wisc.edu>
Content-Type: text/plain; charset="iso-8859-1"
Help!
I don't seem to have recieved the e-mail giving me the site to download the lecture materials. Anyone have it?
Claire
------------------------------
Message: 21
Date: Wed, 28 May 2008 09:33:31 -0500
From: "Juan Gutierrez" <jgutierrez <@t> precisionpath.us>
Subject: RE: [Histonet] heat + formalin
To: "'Nancy Schmitt'" <nancy_schmitt <@t> pa-ucl.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <004201c8c0cf$ce555040$6a00a8c0 <@t> precisionpath.lcl>
Content-Type: text/plain; charset="US-ASCII"
Hi Nancy, if you are doing breast prognostic markers on your cases, you
might want to let it fix overnight before processing. The FDA requires it
for Her-2 testing. No we do not use heat on our formalin steps, but we
require our docs to cut tissue at 3mm or less thickness. So far this has
been working fine with fixing and infiltration.
Good luck,
Juan C. Gutierrez, HT(ASCP)
Precision Pathology Services
210.646.0890
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Nancy
Schmitt
Sent: Wednesday, May 28, 2008 9:09 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] heat + formalin
Good Morning
Need some input, please. Does anyone combine heat during the formalin part
of processing? We are having some issues with breast cases that need to be
read out the next am - we are running them on a shorter processing schedule
(done at 4 am) - and not having the best results.We use a closed system
Tissue-Tek VIP 3000. Is anybody using microwave fixation?
I think we will just put them on the longer processor (done at 0630) and be
done - but wanted to check for other options.
Thanks in advance
Nancy Schmitt
Histology Coordinator
Dubuque, IA
NOTICE: This email may contain legally privileged information. The
information
is for the use of only the intended recipient(s) even if addressed
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------------------------------
Message: 22
Date: Wed, 28 May 2008 07:45:20 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] heat + formalin
To: Nancy Schmitt <nancy_schmitt <@t> pa-ucl.com>,
"'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <743811.30001.qm <@t> web65704.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
I never used heat with the formalin because fumes issues (if by any chance we had to open the retort).
You should not run breast in short cycles unless the slices are extremely thin and you increase the time in the clearing agent (taking away from the alcohols).
It is better to have a good section a few hours later, than a useless section rapidly.
René J.
Nancy Schmitt <nancy_schmitt <@t> pa-ucl.com> wrote:
Good Morning
Need some input, please. Does anyone combine heat during the formalin part
of processing? We are having some issues with breast cases that need to be
read out the next am - we are running them on a shorter processing schedule
(done at 4 am) - and not having the best results.We use a closed system
Tissue-Tek VIP 3000. Is anybody using microwave fixation?
I think we will just put them on the longer processor (done at 0630) and be
done - but wanted to check for other options.
Thanks in advance
Nancy Schmitt
Histology Coordinator
Dubuque, IA
NOTICE: This email may contain legally privileged information. The information
is for the use of only the intended recipient(s) even if addressed
incorrectly. If you are not the intended recipient, please notify the sender
that you have received it in error and then delete it along with any
attachments. Thank you.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 23
Date: Wed, 28 May 2008 11:12:43 -0400
From: stephanie.d.rivera <@t> gsk.com
Subject: [Histonet] Formalin alternative fixative for rodent and large
animal
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<OF131BD0BF.EC8FC68D-ON85257457.0053323E-85257457.00538FFB <@t> gsk.com>
Content-Type: text/plain; charset=us-ascii
Hello histoworld,
Does anyone have any suggestions for altenative fixative to formalin? I
still need to maintain quality of IHC and H&E stain and morphology. I've
come across Prefer and Notoxhisto. Are there any others for animal tissue
that anyone have tried?
Stephanie D. Rivera
Safety Assessment Department
GlaxoSmithKline
709 Swedeland RD
King of Prussia, PA 19406
phone: 610-270-7340
fax: 610-270-7202
------------------------------
Message: 24
Date: Wed, 28 May 2008 11:20:18 -0400
From: "Nicole C Walsh" <nicole.walsh <@t> umassmed.edu>
Subject: [Histonet] Experiencing Chatter in paraffin ribbons
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <94A5E686-2355-422F-8F12-27F149E0EAB8 <@t> umassmed.edu>
Content-Type: text/plain; charset=us-ascii; delsp=yes; format=flowed
Hi,
We routinely cut 5micron paraffin sections of decalcified mouse bones
including whole hindpaws. We use Surgipath Infiltration medium for
infiltration steps and then Surgipath EM-400 Embedding paraffin for
embedding. This has worked well in the past, but since moving to a
new institution and using different microtome setup we are now having
problems with sectioning.
Particularly we are experiencing a lot of chatter in the surrounding
wax. We routinely keep the blocks cold when sectioning.. cooling in
between each ribbon of 5-7 sections (maintained at 4degC on cooling
block on embedding station, with slight coating of block softener
(60% glycerol, 20% ethanol and 20% water)). The tissue generally
comes off intact suggesting that infiltration of the tissue is
reasonable.
We are trying to eliminate possible reasons for our problems.
A couple of things have changed since moving here..
We recently refreshed our paraffin supplies and this seems to have
been when our troubles really started. But Surgipath have said that
they have not changed the formula, but have given us paraffin from a
different lot number to try.
We've had a maintenance rep recently service the microtome (Microm
HM315) and he could not find reason for the chatter (we section with
angle set to 10 as this is what the rep suggested should be used for
this microtome, and have tried varying the angle previously with no
success)
We used to use surgipath high profile blades (non-coated) with
success (and I've seen discussion on histonet to say that the high
profile blades are better for hard tissue).
But the microtome that we now use will only take low-profile blades
and so far we've tried the Richard Allan Scientific low profile
blades (found that these became blunt quickly) and are now trying the
teflon coated low profile blades from surgipath.
If anyone has any suggestions for things that we might try that could
help correct our problems we would be very grateful.
Thanks in advance,
Nicole
------------------------------
Message: 25
Date: Wed, 28 May 2008 10:28:51 -0500
From: "Mahoney,Janice A" <JMahoney <@t> alegent.org>
Subject: [Histonet] RE: NSH teleconference today
To: Ingles Claire <CIngles <@t> uwhealth.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<346E5878979BA54FB4B0BFD6AD93B9B9B01EE96359 <@t> EXCHMBC1.ad.ah.local>
Content-Type: text/plain; charset="us-ascii"
Me either!!!!!
Janice Mahoney HT(ASCP)
Histology/Cytology Coordinator
Alegent Health Laboratory
4955 F Street
Omaha, NE
(402)717-2889
fax(402)717-5231
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ingles Claire [CIngles <@t> uwhealth.org]
Sent: Wednesday, May 28, 2008 9:30 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] NSH teleconference today
Help!
I don't seem to have recieved the e-mail giving me the site to download the lecture materials. Anyone have it?
Claire
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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------------------------------
Message: 26
Date: Wed, 28 May 2008 16:31:11 +0100
From: "Marshall Terry Dr, Consultant Histopathologist"
<Terry.Marshall <@t> rothgen.nhs.uk>
Subject: RE: [Histonet] Rate of formalin penetration in human brain
sections
To: "Geoff McAuliffe" <mcauliff <@t> umdnj.edu>, <Karen_Skish <@t> rush.edu>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<5C0BED61F529364E86309CADEA63FEF20163F40A <@t> TRFT-EX01.xRothGen.nhs.uk>
Content-Type: text/plain; charset="us-ascii"
Agree with Geoff entirely.
In particular you will not find it possible to define an end-point for
formalin fixation.
Terry
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Geoff
McAuliffe
Sent: 28 May 2008 15:19
To: Karen_Skish <@t> rush.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Rate of formalin penetration in human brain
sections
Hi Karen:
Formalin fixes tissue slowly, even very thin slices. This has been
known for many, many years. The work of Medwar and of John R. Baker
(Principles of Biological Technique, John Wiley and Sons, 1958) comes to
mind. Perhaps there are slight differences between kidney and brain but
my guess is that if there is a difference it is insignificant. Before
trying to design (how are you going to define fixation?) and perform
such investigations I suggest a trip to the library. I doubt if the
information you seek is on line, it is too old. However, do not confuse
"old" with "out dated" or "bad". Good luck.
Geoff
Karen_Skish <@t> rush.edu wrote:
> Hi--
> One of our investigators is interested in the approximate rate of
> fixation of human brain tissue, independent of any formaldehyde
diffusion effects.
> In other words, in a very small or very thin piece of human brain
> tissue, what is the fixation rate? He found published data for rat
> kidney, but would like to try to at least determine if the fixation
> rate should be higher or lower in human brain tissue. He is looking
> for data for room temperature, but any information would be greatly
appreciated.
> Thanks!
> Karen M Skish, MS, PA(ASCP)MT
> Pathologists' Assistant & Manager, Neuropathology Lab Rush Alzheimer's
> Disease Center Cohn Research Building, Lab 441
> 1735 West Harrison Street
> Chicago IL 60612
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
--
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
**********************************************
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
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End of Histonet Digest, Vol 54, Issue 41
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