[Histonet] RE: In need of tissue and questions about

Emmanuelle Roux eroux <@t> medimabs.com
Tue Sep 4 13:23:19 CDT 2007


Hi Mindy,
For finding tissues, try Asterand.  They have a lot of human tissues, both
normal and disease states.
Emmanuelle

Emmanuelle Roux, Ph.D.

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Subject: Histonet Digest, Vol 46, Issue 3

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Today's Topics:

   1. RE: damage to dura; also cresyl violet (Kemlo Rogerson)
   2. RE: TUNEL (Bernice Frederick)
   3. PT in a research lab (Helen E Johnson)
   4. RE: Periodic Acid Formalin Fixation (Denise Piontek)
   5. Re: PT in a research lab (Rene J Buesa)
   6. RELIA Histology Job Alert - NYC two positions. (Pam Barker)
   7. In need of tissue and questions about restaining......
      (Mindy Johnson)


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Message: 1
Date: Tue, 4 Sep 2007 11:25:53 +0100
From: "Kemlo Rogerson" <Kemlo.Rogerson <@t> waht.swest.nhs.uk>
Subject: RE: [Histonet] damage to dura; also cresyl violet
To: "John Kiernan" <jkiernan <@t> uwo.ca>, "Caroline Bass"
	<cbass <@t> bidmc.harvard.edu>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<86ADE4EB583CE64799A9924684A0FBBF0222EC4E <@t> wahtntex2.waht.swest.nhs.uk>
Content-Type: text/plain;	charset="us-ascii"

That sounds cruel; hide as there may be animal protectionists around,
they'd blow you up in the UK!!!!

Kemlo Rogerson
Pathology Manager
DD   01934 647057 or extension 3311
Mob 07749 754194; Pager 07659 597107;


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Message: 2
Date: Tue, 4 Sep 2007 08:34:11 -0500
From: "Bernice Frederick" <b-frederick <@t> northwestern.edu>
Subject: RE: [Histonet] TUNEL
To: <koellingr <@t> comcast.net>, "'JR R'" <rosenfeldtek <@t> hotmail.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <006601c7eef8$4a55b720$d00f7ca5 <@t> lurie.northwestern.edu>
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You could probably run a Survivin antibody for comparasion.
Bernice

Bernice Frederick HTL (ASCP)
Northwestern University
Pathology Core Facility
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611
312-503-3723


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
koellingr <@t> comcast.net
Sent: Friday, August 31, 2007 6:17 PM
To: JR R; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] TUNEL

Jerry,
Could you expand on or give references to formalin causing DNA strand
breaks?  Double strand breaks, single strand, blunt end, overhanging?  My
pile of papers and having done TUNEL for years says that formalin fixation
is a very good technique for TUNEL and many peer-reviewed articles in which
TUNEL is used as a technique, use formalin fixation and how can that be if
formalin is causing strand breaks?  In fact one paper I'm looking at says
that extended (5-7 weeks in formalin) fixation causes loss of TUNEL signal.
If formalin is causing breaks, you would assume that TUNEL pos signals would
increase with extended formalin fixation.  Even the use of the monoclonal
antibody F7-26, for single stranded DNA, touts formalin fixation for their
claims of discriminating apoptosis from necrosis.

The question asks about extended alcohol fixation but your answer is
possibly a lot of false positives because formalin causes DNA breaks.  Does
this imply that alcohol won't?  Have done a lot of TUNEL on alcohol fixed
samples.  True I couldn't pretreat them and handle them they way I would
handle FFPE tissue.  Also true that we could argue specificity and ability
or not to discriminate apoptosis from necrosis for quite a while.  But if
formalin itself is causing the breaks in DNA, I and a lot of people are in
big trouble with our science projects and experiments.  Also I can't
envision why 2 cells are showing TUNEL positivity while 2 of the same type
of cells right next to them (and getting the same formalin fix), are
absolutely clean and there is no background?

Thanks for any information you can provide.

Ray Koelling
PhenoPath Laboratories
Seattle, WA

-------------- Original message -------------- 
From: JR R <rosenfeldtek <@t> hotmail.com> 

> 
> I expect you will get a very high background--lots of false positives.
That's a 
> problem with TUNEL and formalin fixed tissue anyway--formalin causes DNA
strand 
> breaks. 
> 
> 
> 
> Jerry L. Ricks 
> Research Scientist 
> U.W. Medicine at South Lake Union 
> 815 Mercer Street 
> Seattle, WA 98109 
> (206)-685-7190> Date: Fri, 31 Aug 2007 10:42:32 +0200> From: 
> marijke.oste <@t> ua.ac.be> To: histonet <@t> lists.utsouthwestern.edu> Subject: 
> [Histonet] TUNEL> > For staining the apoptotic cells I'm already using a
kit of 
> Roche> diagnostics. I think the greatest problem is that my tissues has
been 
> saved> for several years in ethanol 70%. Does anyone has experience in
staining> 
> tissues for apoptotic cells who are kept in ethanol for several years?>
Thank 
> you> > > _______________________________________________> Histonet mailing
list> 
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------------------------------

Message: 3
Date: Tue, 4 Sep 2007 09:44:42 -0400
From: Helen E Johnson <hej01 <@t> health.state.ny.us>
Subject: [Histonet] PT in a research lab
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	
<OF2262886C.94159E9B-ON8525734C.004AD835-8525734C.004B83DF <@t> notes.health.stat
e.ny.us>
	
Content-Type: text/plain; charset=US-ASCII


Hi Histonetters,
      I'm inquiring if there is any Proficiency Testing for research
laboratories.
                                                                    Helen
Johnson  (hej01 <@t> health.state.ny.us)


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Message: 4
Date: Tue, 4 Sep 2007 09:46:06 -0400
From: Denise Piontek <dbpiontek <@t> hotmail.com>
Subject: RE: [Histonet] Periodic Acid Formalin Fixation
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BAY134-W43938EC887D7EA1D6448FAABCA0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"


One last check for info after Labor Day:). Thanks for any advice per the
question below: Dear Histonetters:
 I am performing Periodic Acid Formalin fixation on liver and noticing
sinusoid shrinkage, as well as some edge effect? This is in comparison to a
matched tissue control in 10% NBF. My recipe requires fixation at 4C for 48
hours via 1 gram of periodic acid in 100 mls 10% NBF. Anyone else using this
fixation method? Any suggestions or advice is greatly appreciated,
Denise Bland-Piontek, HTL(ASCP)CTBS(AATB)> NIBRI  
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------------------------------

Message: 5
Date: Tue, 4 Sep 2007 07:19:09 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] PT in a research lab
To: Helen E Johnson <hej01 <@t> health.state.ny.us>,
	histonet <@t> lists.utsouthwestern.edu
Message-ID: <77038.95951.qm <@t> web61213.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Helen:
Proficiency testings always refer to the tasks, not to the place where those
tasks are completed.
  You will have to have the same proficiency to section, regardless of the
setting, although perhaps minor variations could exist.
  Reni J.

Helen E Johnson <hej01 <@t> health.state.ny.us> wrote:
  
Hi Histonetters,
I'm inquiring if there is any Proficiency Testing for research
laboratories.
Helen
Johnson (hej01 <@t> health.state.ny.us)


IMPORTANT NOTICE: This e-mail and any attachments may contain confidential
or sensitive information which is, or may be, legally privileged or
otherwise protected by law from further disclosure. It is intended only for
the addressee. If you received this in error or from someone who was not
authorized to send it to you, please do not distribute, copy or use it or
any attachments. Please notify the sender immediately by reply e-mail and
delete this from your system. Thank you for your cooperation.


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

Message: 6
Date: Tue, 4 Sep 2007 10:37:31 -0400
From: "Pam Barker" <relia1 <@t> earthlink.net>
Subject: [Histonet] RELIA Histology Job Alert - NYC two positions.
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <E1ISZX2-0000PN-EQ <@t> elasmtp-mealy.atl.sa.earthlink.net>
Content-Type: text/plain;	charset="iso-8859-1"

Hi Histonetters,
I am currently working with a client in New York City looking for 2
histo techs.  These are full time permanent positions.  My client offers
excellent compensation, benefits and a prestigious environment to work
in.  For more information please contact me at relia1 <@t> earthlink.net or
toll free at 866-607-3542.  Thanks and 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


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

Message: 7
Date: Tue, 4 Sep 2007 09:50:49 -0700
From: "Mindy Johnson" <m5johnso <@t> meded.ucsd.edu>
Subject: [Histonet] In need of tissue and questions about
	restaining......
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <00dc01c7ef13$bfa028b0$3ee07a10$@ucsd.edu>
Content-Type: text/plain;	charset="US-ASCII"

Hello to all..

 

I am fairly new to histonet.  I am not a histologist by trade, but I am
doing a bunch of histology work at UCSD.  I have a few questions.  Does
anyone know where we can find some pathological tissue?  We need this for
students sets of pathology for our medical students.  It has become pretty
hard to find tissue.  I scoured the internet and found Biomax, but that has
been it.

 

Also, since I have no formal training (I wish I would have taken histology
courses in school) I need some advice on restaining some old tissue.  This
is for our student sets again.  The stain has faded significantly.  Most of
the tissue is at least 15 yrs old.  Some older.   Some a about 10 yrs old.
I have been soaking them in xylene for weeks and I can't seem to 1) either
get the resin off the slide or 2) I have no clue what this option is as they
have been soaking for so long and I do several changes!!  I tried restaining
some of they are pretty much ruined!  So, I have some others and could
really use some help!  If there is any.  Could the tissue be so old and used
so often under the microscope that it has degraded and is not useful
anymore?

 

This is what I have been doing:

 

Soak in xylene (do several changes)

Bleach with potassium permanganate and oxalic acid.

Restain with H&E protocol.

 

But, that wasn't working and was not consistent at all.  So I just tried
putting the slides back in the hematoxylin after soaking in xylene and I got
better results, with one type of tissue, but the older tissue didn't work so
well.

 

Thank you for all your help!!!

 

 Mindy A Johnson

SRA II

UCSD - School of Medicine

Medical Teaching Labs

Fax: 858-822-5250

 



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