[Histonet] RE: Histonet Digest, Vol 48, Issue 30
Pierre CHAUMAT
pierre.chaumat <@t> alphelys.com
Wed Nov 21 12:31:39 CST 2007
Dear Gudrun,
I know using isopentane without a dedicated machine is somehow acrobatic and
difficult. Moreover, maintaining the right temp is also a tricky exercise.
We have developped a system calles SnapFrost that uses electrical
compressors to cool down to -80°C an isopentane bath that will provide you
with ultra-reproducible temperature for always high quality frozens. A path
from Zürich Univ Hosp has two of these units used in routine since a while.
He has shown very good morphology and always much better than freezing bar
of a cryostat. Additionally, it takes only few seconds to get there when a
cryostat bar will take few minutes and time always counts especially in
frozen diagnosis.
Kind regards
Pierre
Your message----------------------------
Message: 5
Date: Wed, 21 Nov 2007 18:50:42 +0100
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: [Histonet] frozens from lung-tissue
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000601c82c67$09d1b0d0$6412a8c0 <@t> dielangs.at>
Content-Type: text/plain; charset="us-ascii"
Hi to all,
I need the help of those, who have experience on making good frozens from
lung-tissue. We look for a method for freezing lung-tissue without the usage
of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the
institute is not satisfied with the quality and wants us to do better. Our
usual method is to make a OCT-layer on a chuck, let it freeze, give the
tissue on it, surround with OCT, put the cold weight on it, let it freeze
and make the sections.
We tried putting the tissue in the liquid OCT on the chuck without a layer
and freezing it in a mold with the chuck on top. I also tried first
freezing the piece solely in liquid nitrogen or isopentan, then putting it
on a chuck. All in all - my boss wasn't happy with it.
Any input will be appreciated. Thank you in advance
Gudrun Lang
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Objet : Histonet Digest, Vol 48, Issue 30
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Today's Topics:
1. Re: Histogel Question (Dana Settembre)
2. RE: How to keep cryostating without getting frostbite?
(Andrea Grantham)
3. RE: Anti-dog IgG (Abigail M. Butler)
4. -80 degree freezer, repair question (Monfils, Paul)
5. frozens from lung-tissue (Gudrun Lang)
----------------------------------------------------------------------
Message: 1
Date: Wed, 21 Nov 2007 11:37:44 -0500
From: "Dana Settembre" <settembr <@t> umdnj.edu>
Subject: Re: [Histonet] Histogel Question
To: <histonet <@t> lists.utsouthwestern.edu>, "Abigail M. Butler"
<am3309 <@t> uga.edu>
Message-ID: <s7441890.034 <@t> smtpnpc.umdnj.edu>
Content-Type: text/plain; charset=US-ASCII
Abbie,
I have used Histogel to embed very small human eye biopsies.
What do you mean, "that obviously is not placed in a tube?"?
We pick up the small specimen carefully with forceps and place the tissue at
the very bottom of the gel and when it goes into a cassette, it is placed
facing down. We make sure that the embedding histotech understands exactly
where it is for processing and embedding.
Dana Settembre, HT ASCP
Immunohistochemistry Lab
UMDNJ - University Hospital
Newark, NJ USA
>>> "Abigail M. Butler" <am3309 <@t> uga.edu> 11/21/07 10:13 AM >>>
I completely understand how to use histogel to embed free floating
cells. But how would I embed a piece of tissue that obviously is not
placed in a tube?
I am considering purchasing Histogel for the first time and have many
ideas of how I could use it, just not sure the proper steps to take to
embed let's say two mice eyes.
Thanks
Abbie Butler, HT (ASCP), QIHC
University of Georgia
College of Veterinary Medicine
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------------------------------
Message: 2
Date: Wed, 21 Nov 2007 10:22:36 -0700
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Subject: RE: [Histonet] How to keep cryostating without getting
frostbite?
To: "Monfils, Paul" <PMonfils <@t> Lifespan.org>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<6.2.3.4.1.20071121100954.01faafa8 <@t> algranth.inbox.email.arizona.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
I cut little rectangular pieces of styrofoam from any of the cartons
that are shipped in with cold or frozen items and "glue" them to each
side of the knife holder with OCT. This provides a great area to rest
my hands while having to manipulate a frozen section. The styrofoam
doesn't get cold and it is a comfty place to put your hands and also
a slide if you have to pick up multiple sections on one slide. The
tissue and OCT doesn't thaw or the slide doesn't get too cold for
another section to attach.
This excellent piece of advice came from Gayle Callis - thank you
again Gayle!!!
PS: When I want to clean out the cryostat I just turn it off and take
out the knife holder and when the OCT warms up the styrofoam comes
off and can be reattached when the cryostat is all clean and ready to
be turned back on.
I was reading where somebody suggested something to a manufacturer
and the idea was not readily accepted and I likewise have made what I
thought were excellent suggestions and was met with similar responses
- this styrofoam thing was just one of them.
Andi
At 09:16 AM 11/21/2007, Monfils, Paul wrote:
>Just having your hands in the cold air isn't usually the problem.
>Each tech, according to their particular style, usually rests their
>hand on the knife holder in some manner for stability while picking
>up the sections on the slides. The rate of extraction of heat by
>contact with -20 degree metal is much greater than the rate by
>simple exposure to -20 degree air. It is these points of repeated
>contact between skin and metal that usually suffer from
>frostbite. I have tried various kinds of gloves and slip-on finger
>protectors, but in the final analysis I find that bandaids carefully
>applied to the specific points of contact provide enough insulation
>to prevent skin damage. I buy the extra wide ones.
>
> > ----------
> > From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf
> of jstaruk
> > Sent: Tuesday, November 20, 2007 7:49 PM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] How to keep cryostating without getting
frostbite?
> >
> > Hey all,
> >
> > Are there any tricks on how to keep your fingers from getting frostbite
> > while doing cryostat sectioning for 8 hours a day? My poor techs who
have
> > been cutting frozen sections for several straight days now are starting
to
> > complain about these inhumane conditions! Do all of you MOHS techs have
> > bleeding, chapped hands and numb fingertips?
> >
> > Thank you in advance for any suggestions and Happy Thanksgiving to all!
> >
> > Jim
> >
> > _____________________
> > Jim Staruk
> > Mass Histology Service
> > www.masshistology.com
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
>_______________________________________________
>Histonet mailing list
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.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist University of Arizona :
: (office: AHSC 4212) P.O. Box 245044 :
: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
------------------------------
Message: 3
Date: Wed, 21 Nov 2007 12:28:14 -0500 (EST)
From: "Abigail M. Butler" <am3309 <@t> uga.edu>
Subject: [Histonet] RE: Anti-dog IgG
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20071121122814.JJF06401 <@t> punts1.cc.uga.edu>
Content-Type: text/plain; charset=us-ascii
MaryAnn,
I use only anti-dog IgG.....and my dilution is 1:150,000.
Crazy, isn't it? I also used HIER for pretreatment with citrate buffer.
There is not much background.
Hope this helps
Abbie Butler, HT (ASCP), QIHC
University of Georgia
College of Veterinary Medicine
------------------------------
Message: 4
Date: Wed, 21 Nov 2007 12:36:05 -0500
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: [Histonet] -80 degree freezer, repair question
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4EBFF65383B74D49995298C4976D1D5E273CFC <@t> LSRIEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain; charset="iso-8859-1"
We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree
freezer that works fine except the lid gasket is shot, so cold air leaks out
and frost and ice build up around the edges of the lid. Does anyone know of
someone who could replace this gasket? Or somewhere we could purchase the
part to install ourselves? We are also unable to find particle filters for
the front door of the compressor compartment.
------------------------------
Message: 5
Date: Wed, 21 Nov 2007 18:50:42 +0100
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: [Histonet] frozens from lung-tissue
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000601c82c67$09d1b0d0$6412a8c0 <@t> dielangs.at>
Content-Type: text/plain; charset="us-ascii"
Hi to all,
I need the help of those, who have experience on making good frozens from
lung-tissue. We look for a method for freezing lung-tissue without the usage
of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the
institute is not satisfied with the quality and wants us to do better. Our
usual method is to make a OCT-layer on a chuck, let it freeze, give the
tissue on it, surround with OCT, put the cold weight on it, let it freeze
and make the sections.
We tried putting the tissue in the liquid OCT on the chuck without a layer
and freezing it in a mold with the chuck on top. I also tried first
freezing the piece solely in liquid nitrogen or isopentan, then putting it
on a chuck. All in all - my boss wasn't happy with it.
Any input will be appreciated. Thank you in advance
Gudrun Lang
------------------------------
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