[Histonet] RE: Frozen Section Help
Biedermann, JoAnn
JABiedermann <@t> uams.edu
Mon Nov 8 14:10:49 CST 2010
Did you cryoprotect the tissue before you cut it?
JB
Jo Ann Biedermann
Research Assistant
University of Arkansas for Medical Sciences
Reynolds Institute on Aging
629 Jack Stephens Drive
Room 3173 Mail Slot 807
Little Rock, AR 72205
Phone: 501-526-5803
FAX: 501-526-5830
JABiedermann <@t> uams.edu
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Today's Topics:
1. cytotechnology (Mimmo Pozzuoli)
2. RE: SPAM-LOW: [Histonet] shrinkage during IHC (Patsy Ruegg)
3. RE: SPAM-LOW: [Histonet] cytotechnology (Patsy Ruegg)
4. Frozen section ...HELP (louise renton)
5. Re: Frozen section ...HELP (Lee & Peggy Wenk)
6. RE: shrinkage during IHC (Kuhnla, Melissa)
7. RE: Frozen section ...HELP (Monfils, Paul)
8. Distance Learning (Phyllis Thaxton)
9. Microwave processing effect on DNA/RNA (Matt Brooks)
----------------------------------------------------------------------
Message: 1
Date: Sun, 7 Nov 2010 13:36:09 -0500
From: Mimmo Pozzuoli <mimmopozzuoli <@t> gmail.com>
Subject: [Histonet] cytotechnology
To: histonet <@t> lists.utsouthwestern.edu
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<AANLkTim4E0D0+LOFA=WtkCprr+THeP7H3ew7-B=tK7aW <@t> mail.gmail.com>
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Are any histotechs out there performing FISH or automated cyto procedures in
their labs? If so, are your pathologists having to screen and act as the
cyto supervisors?
What are the guidelines on this? Can a histotech perform cytoprep functions,
or is the slide screening integral to the position?
------------------------------
Message: 2
Date: Sun, 7 Nov 2010 13:20:58 -0700
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: SPAM-LOW: [Histonet] shrinkage during IHC
To: <zodiac29 <@t> comcast.net>, <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <9109304A049D40F889C150360425E41B <@t> prueggihctechlt>
Content-Type: text/plain; charset="us-ascii"
You should have the same morphology from the IHC slides done outside that
you get from your H&E, do you provide them with slides or the Block? The
only thing I can think of is if they airdry after IHC instead of going thru
alcohols and xylene to coverslip (if they use AEC or ap/red they may
airdry), I have seen cell shrinkage from airdrying after IHC occasionally.
Something else just occurred to me, if they over heat during HIER I suppose
this could happen, but I have not seen it.
Regards,
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net
www.ihcrg.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
zodiac29 <@t> comcast.net
Sent: Saturday, November 06, 2010 6:28 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] shrinkage during IHC
To All,
We are a lab that sends our specimens out for IHC and have just switched to
another reference laboratory for these services. Our pathologist is saying
that the tissue looks shrunk on the IHC slides, yet the slides that I
process (H&E, and special stains)are fine. Does anyone know what is causing
this? The reference lab said it could be the type of slides that I use to
mount the sections we send to them. My knowledge in IHC is limited. Also, if
this helps, they are FFPE tissue.
Thanks for your help
Jenny
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------------------------------
Message: 3
Date: Sun, 7 Nov 2010 13:55:42 -0700
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: RE: SPAM-LOW: [Histonet] cytotechnology
To: "'Mimmo Pozzuoli'" <mimmopozzuoli <@t> gmail.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <FCFB1EBA62D341D29251230913EB0BE0 <@t> prueggihctechlt>
Content-Type: text/plain; charset="us-ascii"
Are you talking about interpreting the test or just performing it?
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net
www.ihcrg.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mimmo
Pozzuoli
Sent: Sunday, November 07, 2010 11:36 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] cytotechnology
Are any histotechs out there performing FISH or automated cyto procedures in
their labs? If so, are your pathologists having to screen and act as the
cyto supervisors?
What are the guidelines on this? Can a histotech perform cytoprep functions,
or is the slide screening integral to the position?
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 4
Date: Mon, 8 Nov 2010 11:43:13 +0200
From: louise renton <louise.renton <@t> gmail.com>
Subject: [Histonet] Frozen section ...HELP
To: Histonet <Histonet <@t> lists.utsouthwestern.edu>
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<AANLkTimsXAMzcyVmX+8w6RHgZ0uuBemBMSt-1=kVhsDT <@t> mail.gmail.com>
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Hi all,
after more than a decade of NOT cutting frozen sections, I find myself
back at the ice-face.
To get my hand in (not literally) I thought I would do some trial
sections on stored tissue - stuff that was in formalin and now in 70%
alcohol.
Horror ; dismay. The tissue, once frozen, is all mushy in the middle.
Is this because of teh long storage?
is there anything I can do to improve the situation?
much appreciated
--
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
+27 11 717 2298 (tel & fax)
073 5574456 (emergencies only)
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
------------------------------
Message: 5
Date: Mon, 8 Nov 2010 05:33:52 -0500
From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
Subject: Re: [Histonet] Frozen section ...HELP
To: "louise renton" <louise.renton <@t> gmail.com>, "Histonet"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <0B2A6F65620A4565904BC1C81A1B76F9 <@t> HP2010>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=original
The freezing point of water is 0 degrees C. The freezing point of 100%
ethanol is -114 degrees C. The freezing point of 70% alcohol is about -48
degrees C. Since most cryostats are at -20 to -25 degrees C, your tissue
isn't freezing completely. You probably have "slush" ice inside the cells.
Not hard enough to support the tissue during a frozen section.
Fresh tissue would be the best, as formalin fixed tissue also tends to cut
awful (whole different reason). If you can't get fresh tissue from human or
animal necropsy, can you get some "parts" from a raw uncooked chicken to
practice on? Save some muscle, skin, liver, etc., from tonight's supper?
Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073
--------------------------------------------------
From: "louise renton" <louise.renton <@t> gmail.com>
Sent: Monday, November 08, 2010 4:43 AM
To: "Histonet" <Histonet <@t> lists.utsouthwestern.edu>
Subject: [Histonet] Frozen section ...HELP
> Hi all,
>
> after more than a decade of NOT cutting frozen sections, I find myself
> back at the ice-face.
> To get my hand in (not literally) I thought I would do some trial
> sections on stored tissue - stuff that was in formalin and now in 70%
> alcohol.
>
> Horror ; dismay. The tissue, once frozen, is all mushy in the middle.
> Is this because of teh long storage?
>
> is there anything I can do to improve the situation?
>
> much appreciated
>
> --
> Louise Renton
> Bone Research Unit
> University of the Witwatersrand
> Johannesburg
> South Africa
> +27 11 717 2298 (tel & fax)
> 073 5574456 (emergencies only)
> "There are nights when the wolves are silent and only the moon howls".
> George Carlin
> No trees were killed in the sending of this message.
> However, many electrons were terribly inconvenienced.
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Mon, 8 Nov 2010 08:08:25 -0500
From: "Kuhnla, Melissa" <Melissa.Kuhnla <@t> chsli.org>
Subject: RE: [Histonet] shrinkage during IHC
To: <zodiac29 <@t> comcast.net>, <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<C76F12086768614F9C2618ED9966FEE10698F392 <@t> MMDCNT0BXVS003.chsli.org>
Content-Type: text/plain; charset="us-ascii"
In my experience, IHC usually plumps tissue back up during reteival. Is
this noticed with every antibody or just a few? Certain tissue types?
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
zodiac29 <@t> comcast.net
Sent: Saturday, November 06, 2010 8:28 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] shrinkage during IHC
To All,
We are a lab that sends our specimens out for IHC and have just switched
to another reference laboratory for these services. Our pathologist is
saying that the tissue looks shrunk on the IHC slides, yet the slides
that I process (H&E, and special stains)are fine. Does anyone know what
is causing this? The reference lab said it could be the type of slides
that I use to mount the sections we send to them. My knowledge in IHC is
limited. Also, if this helps, they are FFPE tissue.
Thanks for your help
Jenny
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Message: 7
Date: Mon, 8 Nov 2010 09:14:09 -0500
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] Frozen section ...HELP
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4EBFF65383B74D49995298C4976D1D5E075E13A2 <@t> LSRIEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain; charset="us-ascii"
Hopefully you removed the antifreeze (alcohol) before freezing? :-) I
have run into this a few times. I work in a core facility where people
send me samples from all over. Recently someone sent me some samples
fixed in Histofix, which is a commercially available aqueous fixative.
But they neglected to tell me they add 15% ethanol to the commercial
solution. I froze all the samples in OCT compound, but they could not
be sectioned. I had to melt them all, soak them in several changes of
buffer for several hours to remove the OCT and the methanol, then put
them in fresh OCT and refreeze them. And that was only 15% alcohol, not
70%!
------------------------------
Message: 8
Date: Mon, 8 Nov 2010 07:26:59 -0800 (PST)
From: Phyllis Thaxton <dchihc <@t> yahoo.com>
Subject: [Histonet] Distance Learning
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <718649.88511.qm <@t> web43512.mail.sp1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
We are thinking of?using one of the online or distance learning programs to help
in the training of new histotechs. Has anyone had any experience with any of
these programs,?what colleges offer the program, pros, cons. Any feedback is
welcome.
Thanks
?Phyllis Thaxton HT(ASCP)QIHC
DCH Regional Medical Center
Tuscaloosa, AL
------------------------------
Message: 9
Date: Mon, 8 Nov 2010 08:36:04 -0800
From: "Matt Brooks" <mbrooks <@t> incytepathology.com>
Subject: [Histonet] Microwave processing effect on DNA/RNA
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<706224670091FE47997AEF88EFADE7CA0194EA35 <@t> EXCHANGE-SRV.PAI.E-PATHOLOGY.COM>
Content-Type: text/plain; charset="us-ascii"
Hello All,
I have seen some posts on the "possible" damaging effects on DNA and RNA
by microwave processing. Does anyone have a reference article that they
can share with me? We are in the process of budgeting for next year and
we are looking at microwave processors; but I want to verify if this
claim is or is not valid.
Thank you,
Matt Brooks, BS, HT (ASCP)
Histology Supervisor
InCyte Pathology
mbrooks <@t> incytepathology.com
509-892-2744
------------------------------
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