[Histonet] RE: bonw marrow mush
Emerson, Rachael
Rachael_Emerson <@t> URMC.Rochester.edu
Thu Nov 4 12:14:07 CST 2004
Hello, I do not think that bone marrow frozen at 4dc straight from
dissection
is going to have any viability. I would refer to Stem Cell Technologies
website.
The have kits and reagents, as well as, great manuals for this kind of
stuff.
Rachael Emerson
Rachael L. Emerson
Center for Human Genetics and Molecular Pediatric Diseases
University of Rochester Medical Center
575 Elmwood Avenue MRBX 1.11301
Rochester, NY 14642
Tel (585) 275-5073
Fax (585) 276-0232
> ----------
> From: histonet-request <@t> lists.utsouthwestern.edu
> Reply To: histonet <@t> lists.utsouthwestern.edu
> Sent: Thursday, November 4, 2004 1:04 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: Histonet Digest, Vol 12, Issue 5
>
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> Today's Topics:
>
> 1. CD41, GPV, GP1bBeta (Emerson, Rachael)
> 2. TB block (Elizabeth Chlipala)
> 3. Fixation for carbohydrates (Hernan Aldana)
> 4. practical exam... (JCarpenter764 <@t> aol.com)
> 5. exam (maryjohnson)
> 6. Using expired reagents in IHC (Amos Brooks)
> 7. Re: Exam (Gareth Davis)
> 8. test (Denise Bland-Piontek)
> 9. Re: Re: Exam (Dndsomi <@t> aol.com)
> 10. EGFR (CrochiereSteve <@t> aol.com)
> 11. Timm silver sulphide method for light microscopic
> localisation of heavy metals. (Chris.Goodall <@t> bristol.ac.uk)
> 12. Histonet 2005 Buttons (ajennings <@t> unmc.edu)
> 13. RE: Job Openings- Texarkana, TX (mprice26 <@t> juno.com)
> 14. Re: Timm silver sulphide method for light
> microscopiclocalisation of heavy metals. (John Kiernan)
> 15. RE: exam (Patsy Ruegg)
> 16. RE: BMP7 (Patsy Ruegg)
> 17. bone marrow mush (Patsy Ruegg)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Wed, 3 Nov 2004 14:00:53 -0500
> From: "Emerson, Rachael" <Rachael_Emerson <@t> URMC.Rochester.edu>
> Subject: [Histonet] CD41, GPV, GP1bBeta
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <CDF18E16753BD61199540003470A8722073C1159 <@t> exms5.urmc.rochester.edu>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi. Does anyone have any experience working with CD41, GPV, or GP1bBeta
> antibodies?
> I could really use some help.
>
> Thanks a lot!!
> Rachael Emerson
>
> Rachael L. Emerson
> Center for Human Genetics and Molecular Pediatric Diseases
> University of Rochester Medical Center
> 575 Elmwood Avenue MRBX 1.11301
> Rochester, NY 14642
>
> Tel (585) 275-5073
> Fax (585) 276-0232
>
>
> > ----------
> > From: histonet-request <@t> lists.utsouthwestern.edu
> > Reply To: histonet <@t> lists.utsouthwestern.edu
> > Sent: Wednesday, November 3, 2004 1:03 PM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: Histonet Digest, Vol 12, Issue 4
> >
> > Send Histonet mailing list submissions to
> > histonet <@t> lists.utsouthwestern.edu
> >
> > To subscribe or unsubscribe via the World Wide Web, visit
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> > or, via email, send a message with subject or body 'help' to
> > histonet-request <@t> lists.utsouthwestern.edu
> >
> > You can reach the person managing the list at
> > histonet-owner <@t> lists.utsouthwestern.edu
> >
> > When replying, please edit your Subject line so it is more specific
> > than "Re: Contents of Histonet digest..."
> >
> >
> > Today's Topics:
> >
> > 1. Alcian Blue and steamer containers (Paula Pierce)
> > 2. Re: Sta-On (Jennifer Sipes)
> > 3. Epidermal growth factor (Ford, Rhonda)
> > 4. Job posting - Dallas (Patterson, Pat)
> > 5. RE: thanks and aquamount (Patsy Ruegg)
> > 6. RE: AFB Stain (Joe Nocito)
> > 7. Re: History of microtechnique (John Kiernan)
> > 8. Re: GMA sectioning (Caroline Stott)
> > 9. Re: sample preparation/analysis to observe gas bubbles
> > (Wayne Kreider)
> > 10. Re: Polyoma Virus (tdobersztyn <@t> chmca.org)
> > 11. Re: AFB Stain (lpwenk <@t> sbcglobal.net)
> > 12. Merkel cell staining (Marshall)
> > 13. LR White vs LR Gold (Peter Rippstein)
> > 14. RE: Modified Gomori's Trichrome (Sharon Allen)
> > 15. Envision System with Vector Red Background (Amy Kozer)
> > 16. Re: Merkel cell staining (Richard Cartun)
> > 17. B & D steamer (Rita Angel)
> > 18. Polyoma BK virus controls (Histology SLU)
> > 19. BMP7 (Grant, Debra)
> > 20. Thanks for the Teasing (Ant S.)
> >
> >
> > ----------------------------------------------------------------------
> >
> > Message: 1
> > Date: Tue, 2 Nov 2004 10:06:53 -0800 (PST)
> > From: Paula Pierce <contact <@t> excaliburpathology.com>
> > Subject: [Histonet] Alcian Blue and steamer containers
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <20041102180653.38389.qmail <@t> web50308.mail.yahoo.com>
> > Content-Type: text/plain; charset=us-ascii
> >
> > Hello Andrew,
> >
> > yes I have already voted today :)
> >
> > You can buy a smaller amount of alcian blue at
> > http://www.anatechltdusa.com/Catalog/catalogspecialstains.html
> >
> > I use plastic coplin jars in my B&D steamer for small quantities of
> slides
> > and the larger tissue-tek when I have more.
> >
> >
> > Paula Pierce, HTL(ASCP)HT
> >
> > Excalibur Pathology, Inc.
> > 631 N. Broadway
> > Moore, OK 73160
> > 405-759-3953
> > contact <@t> excaliburpathology.com
> > www.excaliburpathology.com
> >
> > ------------------------------
> >
> > Message: 2
> > Date: Tue, 2 Nov 2004 10:10:48 -0800 (PST)
> > From: Jennifer Sipes <jengirl1014 <@t> yahoo.com>
> > Subject: [Histonet] Re: Sta-On
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <20041102181048.98641.qmail <@t> web60604.mail.yahoo.com>
> > Content-Type: text/plain; charset=us-ascii
> >
> > I purchased it through a company called Surgi-Path. They're the ones
> that
> > make it.
> >
> >
> > Jennifer K. Sipes, RALAT
> > Sr. Laboratory Technician
> > Johns Hopkins University
> > Ross 929
> > 720 Rutland Avenue
> > Baltimore, MD 21205
> > phone: 410-614-0131
> > cell: 443-413-0853
> > e-mail: jengirl1014 <@t> yahoo.com
> >
> >
> >
> >
> >
> >
> >
> >
> >
> >
> >
> > ---------------------------------
> > Do you Yahoo!?
> > Check out the new Yahoo! Front Page. www.yahoo.com/a
> >
> > ------------------------------
> >
> > Message: 3
> > Date: Tue, 2 Nov 2004 13:11:38 -0500
> > From: "Ford, Rhonda" <RFORD <@t> HCMHCARES.ORG>
> > Subject: [Histonet] Epidermal growth factor
> > To: <histonet <@t> pathology.swmed.edu>
> > Message-ID:
> > <684DF8CDE1FB6A428499DA65D753D12201F61789 <@t> JUPITER.HCMH.ORG>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Does anyone know any institution(s) who perform this testing?
> >
> >
> > ------------------------------
> >
> > Message: 4
> > Date: Tue, 2 Nov 2004 12:26:35 -0600
> > From: "Patterson, Pat" <PatPatterson <@t> mhd.com>
> > Subject: [Histonet] Job posting - Dallas
> > To: "'histonet <@t> lists.utsouthwestern.edu'"
> > <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43B0D <@t> omega.mhd.com>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> >
> > Named one of the "Best Places to work 2004" by the Dallas Business
> > Journal.
> >
> > Our dedicated team is very outspoken in their appreciation of the
> > excellent
> > clinical environment we foster and the ongoing commitment Methodist has
> > made
> > to patient satisfaction. Our dual focus on out patients and our
> employees
> > is working - 2002 Methodist Health System earned the Gold Seal of
> > Approval
> > form the Joint Commission on Accreditation of Healthcare Organizations
> and
> > more than 28 percent of our new hires come from employee referral! If
> > excellent patient care is one of your priorities for career
> satisfaction,
> > then you belong at Methodist.
> >
> >
> >
> > Join our Histology team at Methodist Dallas Medical Center. We're
> seeking
> > a
> > full-time Histology Technician on the day shift in our busy Anatomic
> > Pathology Section. Job responsibilities include processing, embedding,
> > sectioning, routine and special staining, assisting in gross room,
> frozen
> > sectioning and computer entry into the MediTech lab information system.
> > Experience with microwave technology would be an added plus. Requires
> > HT/HTL
> > ASCP registry or eligible. Minimum 2-year experience desired.
> >
> > Contact: Pat Patterson, Manager
> > Phone: (214) 947-3538
> > Fax: (214) 947-3524
> > email: PatPatterson <@t> mhd.com
> >
> >
> > ***********************************************************************
> >
> > This electronic transmission contains information from Methodist Health
> > System and should be considered confidential and privileged. The
> > information contained in the above messages is intended only for the
> > use of the individual(s) and entity(ies) named above. If you are not
> > the intended recipient, be aware that any disclosure, copying,
> > distribution, or use of this information is prohibited. If you receive
> > this transmission in error, please notify the sender immediately by
> > return e-mail. Methodist Health System, its subsidiaries and
> > affiliates hereby claim all applicable privileges related to the
> > transmission of this communication.
> >
> >
> >
> > ------------------------------
> >
> > Message: 5
> > Date: Tue, 2 Nov 2004 11:33:43 -0700
> > From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> > Subject: RE: [Histonet] thanks and aquamount
> > To: "'Sharon Cooperman'" <scoop <@t> mail.nih.gov>,
> > <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <001501c4c10a$7b4e4fa0$83020a0a <@t> IHCTech>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Sharon,
> > Aquamount will not harden, you should seal with clear nail polish to
> > keep it permanently.
> > Patsy
> >
> > -----Original Message-----
> > From: histonet-bounces <@t> lists.utsouthwestern.edu
> > [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sharon
> > Cooperman
> > Sent: Thursday, October 28, 2004 12:16 PM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] thanks and aquamount
> >
> >
> > Dear Histonetters,
> >
> > Thanks to everyone for the great advice on bone decal and oil red o
> > stain. I have one last question related to the oil red o stain - I
> > just bought a bottle of aquamount to use when mounting the oil red o
> > stained slides, but it didn't come with instructions (on the bottle
> > it refers to instructions). Are there any special instructions for
> > aquamount? Do I need to seal the slides with nail polish or does
> > aquamount harden? Or, does anyone know how I can contact Lerner to
> > get a copy of the instructions (I bought it from Fisher - Lerner
> > doesn't answer the phone number I have for them).
> >
> > Thanks,
> > Sharon
> > --
> > Sharon Cooperman <scoop <@t> mail.nih.gov>
> > NIH, NICHD, CBMB 301.435-8417
> > Building 18T, room 101 301.402-0078 fax
> > Bethesda, MD 20892
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 6
> > Date: Tue, 2 Nov 2004 12:58:15 -0600
> > From: "Joe Nocito" <JNocito <@t> Pathreflab.com>
> > Subject: RE: [Histonet] AFB Stain
> > To: "Etheridge, Sandra AGF:EX" <Sandra.Etheridge <@t> gems8.gov.bc.ca>,
> > <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <JFEMICGBHEGPLAMIJPJPIEDHCHAA.JNocito <@t> Pathreflab.com>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> > Sandra,
> > attached is our procedure for Truant's AFB. We purchase the staining kit
> > from Infolab.
> >
> >
> >
> > -----Original Message-----
> > From: histonet-bounces <@t> lists.utsouthwestern.edu
> > [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of
> > Etheridge, Sandra AGF:EX
> > Sent: Tuesday, November 02, 2004 11:57 AM
> > To: (histonet <@t> lists.utsouthwestern.edu)
> > Subject: [Histonet] AFB Stain
> >
> >
> > Hello, all,
> >
> > Does anyone have a current method for acid fast bacilli using
> > auramine-rhodamine? Is this a fluorescent stain?
> >
> > Thanks in advance.
> >
> > Sandra Etheridge
> >
> > BC Ministry of Agriculture, Food & Fisheries
> > Animal Health Center, Histology
> > Abbotsford, BC
> > Canada
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> > ------------------------------
> >
> > Message: 7
> > Date: Tue, 02 Nov 2004 14:19:49 -0500
> > From: John Kiernan <jkiernan <@t> uwo.ca>
> > Subject: Re: [Histonet] History of microtechnique
> > To: MTitford <@t> aol.com
> > Cc: Histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <4187DDD5.6F8C39E1 <@t> uwo.ca>
> > Content-Type: text/plain; charset=us-ascii
> >
> > There is also a chapter on the History of Staining
> > by F. Kasten in the 10th edition of "Conn's
> > Biological Stains." This book (2002) is
> > still in print.
> > -------------------------------
> > John A. Kiernan
> > Department of Anatomy and Cell Biology
> > The University of Western Ontario
> > London, Canada N6A 5C1
> > _______________________________
> > MTitford <@t> aol.com wrote:
> > >
> > > David Muskett asks about where to find information about the history
> of
> > microtechnique.
> > > A good place to start is with "A history of microtechnique" by Brian
> > Bracegirdle. The second edition is still available here in the States
> from
> > Science Heritage Limited, Lincolnwood, IL
> > > a second good book is "History of staining" by George Clark and
> > Frederick Kasten. 3ed edition, Williams & Wilkins, Baltimore & London.
> The
> > second book is probably out of print but available on interlibrary loan.
> > > There are many other sources but you can get information overload in
> > this area!
> > >
> > > Mike Titford
> > > USA Pathology
> > > Mobile AL
> >
> >
> >
> > ------------------------------
> >
> > Message: 8
> > Date: Wed, 03 Nov 2004 09:49:23 +1300
> > From: Caroline Stott <caroline.stott <@t> anatomy.otago.ac.nz>
> > Subject: [Histonet] Re: GMA sectioning
> > To: Histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <5.2.1.1.0.20041103094227.02076a10 <@t> anatomy.otago.ac.nz>
> > Content-Type: text/plain; charset="us-ascii"; format=flowed
> >
> > Hi Ben,
> > We have quite regularly cut GMA at 10, 20, 30 and 40 microns. We use a
> > cotton bud to apply a little water to the face of the block, just gently
>
> > rubbing it in. Also place some water on the glass knife as the section
> > will come off easier. Then take a section and push it to the bottom of
> > the
> > water bowl/dish so it will flatten out. Then slide the sections onto
> the
> > slide. Heres the tricky part.... We put a piece of filter paper on the
>
> > bench then the slide on top, then another piece of wet filter paper on
> top
> >
> > of the slide (and sections) and use a roller (like a paint roller)
> > applying a little pressure while rolling around 20-30 times over the
> slide
> >
> > and finally place on a hotplate (not flat) that is around 60 degrees
> > C. Haven't had a problem. :)
> > Hope that makes sense.
> >
> > Caroline
> >
> > Caroline Stott
> >
> > Histology Service Unit
> > University of Otago
> > PO Box 913
> > Dunedin, New Zealand
> > Ph (03) 479 7152
> > Fax (03) 479 7136
> >
> > ------------------------------
> >
> > Message: 9
> > Date: Tue, 02 Nov 2004 12:54:20 -0800
> > From: Wayne Kreider <wkreider <@t> u.washington.edu>
> > Subject: Re: [Histonet] sample preparation/analysis to observe gas
> > bubbles
> > To: Philip Oshel <peoshel <@t> wisc.edu>
> > Cc: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <4187F3FC.3060400 <@t> u.washington.edu>
> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed
> >
> > Phil,
> > Thanks again... all very helpful. I think we can now plan how we'll
> > have the best chance at 'catching' transient bubbles. I'm not sure
> > we'll have our techniques together for sampling and freezing the tissue
> > in the very near term (this week), but I am hopeful we can do/try this.
>
> > By the way, you mentioned that frozen samples are not useful for EM...
> > how so? Perhaps due to subsequent fixing procedures? I'm relatively
> > unfamiliar with EM although TEM had been suggested for this work.
> > Wayne
> >
> > Philip Oshel wrote:
> >
> > > Wayne,
> > >
> > > You do have a problem ...
> > > 2 X 2 X 2 mm is too big for any proper freezing method. Even
> > > high-pressure freezing, which can in ideal conditions give the
> > > greatest depth of good freezing, only does at best 500 microns. The
> > > other methods do 200 microns or less. Plunging into LN2-cooled
> > > cryogens will do a few 10s of microns, no more. They're fine for light
>
> > > microscopy (assuming migration of molecules from holed membranes or
> > > desiccation isn't an issue), but not EM.
> > > The major source of freezing damage isn't really mechanical
> > > ice-crystal growth and damage, but dehydration of cells from crystal
> > > growth.
> > > Plunging is literally dropping the sample into the cryogen, but not
> > > really. By then I mean you *rapidly* plunge it in, not just let it
> > > fall. By hand, as fast as possible. This is important in order to get
> > > through the layer of cold nitrogen above the cryogen (whatever it is).
>
> > > Some commercial plungers are designed to drop the sample into the
> > > cryogen from enough height to pass through the cold gas layer quickly.
>
> > > These are used for electrophysiology -- stimulate the sample then
> > > drop, or stimulate during the drop, and therefore the sample is frozen
>
> > > X msec after stimulation.
> > > I can see doing this with ultrasonication, depending on the size, etc.
>
> > > of the sonciation apparatus.
> > > Cryoprotectants. Um. I don't think you can use these. The tissue has
> > > to soak in the cryoprotectant -- often moving up a gradient -- in
> > > order to get the cryoprotectant completely infiltrated into the
> > > sample. This takes an hour or more -- maybe several hours? You'd
> > > either have to assume nothing happens with the bubbles in the tissue
> > > while sitting around infiltrating with cryoprotectant, or you'd have
> > > to get the cryoprotectant into the tissue first, then sonicate and
> > > pretend the cryoprotectant doesn't change the tissue properties and
> > > therefore resulting bubbles.
> > > I wouldn't believe either of those choices.
> > > I think you'll need to do the experiment, then freeze, and use pieces
> > > of tissue small enough to freeze properly. But! It's not entirely bad
> > > -- only one dimension has to be =<200 microns -- 100 microns, really.
> > > The other dimensions could be 2 mm.
> > > So: 2 mm X 2 mm X 100 microns (even thinner would be better) should
> > > work. Just hold the tissue by a corner, so that the cryogen has
> > > maximal access to both faces of the thin side. And move the tissue
> > > once in the slush LN2, don't hold it in one place -- keep fresh
> > > cryogen contacting the sample.
> > > Let's see -- oh. Insulate the container (and vacuum desiccator) as
> > > best as possible, and work quickly, since you'll only have a few
> > > minutes to freeze before the slush LN2 warms up to regular
> > > almost-boiling LN2.
> > > (I think Bal-Tec makes an apparatus for making slush LN2, and maybe
> > > freezing with it, but you don't really need it, it just maybe makes
> > > things easier.)
> > >
> > > Phil
> > >
> > > Phil,
> > > Thanks for the input. Unfortunately, I haven't had much luck looking
> > > in the archive.
> > > The treatment is actually acoustic (high intensity ultrasound), so no
> > > electrodes are involved. Our treatment volume that we'd ideally like
> > > to freeze is about 2mm x 2mm x 5mm. However, from the little I've
> > > read about plunging, a guideline for the maximum sample thickness is
> > > 0.2mm. Using a slush as you describe would presumably help some, but
> > > I suspect 2mm is still too thick.
> > > If we are able to obtain a suitably thin sample, is plunging literally
>
> > > just dropping the raw tissue into the cryogen? Or are there some
> > > additional steps that should be taken in handling the sample?
> > > Moreover, if we do have a larger sample, reference books suggest that
> > > it's necessary to use a cryo-protectant. Is a cryo-protectant likely
> > > to distort the tissue, in particular any bubble-type structures?
> > > Also, what time delays are typically associated with using a
> > > protectant? Any suggestions about which cryo-protectant might be best
>
> > > suited for this application? Thanks again... I'd appreciate any
> > > thoughts you might have.
> > > Wayne
> > >
> > > Philip Oshel wrote:
> > >
> > > Cryofixation is the only way I know to preserve this kind of
> > > structure. There have been discussions about this in the past, so you
> > > might want to poke around in the Histonet archives.
> > > What kind of treatment? Is it something that you stimulate with say an
>
> > > electrode?
> > > If so, there are plunge freezers designed to allow you to stimulate
> > > the sample while it is falling into the liquid nitrogen.
> > > Rapidity of freezing is essential to insure vitrification of the
> > > water, instead of crystal formation. Although some folks maintain that
>
> > > the water doesn't vitrify, but instead forms micro-crystallites small
> > > enough to have no effect on structure. Others state that evanescent
> > > microspherules are formed, sort of neither fish nor fowl -- not
> > > crystalline and not glass. Either way, the frozen samples have to be
> > > maintained below the recrystallization temperature (around -80deg C or
>
> > > so) until after dehydraton or embedding to prevent crystal growth and
> > > negating all the rapid freezing goodness.
> > > But, the 4 basic methods are high-pressure freezing, propane-jet
> > > freezing, slam-freezing, and plunging into cryogen. The first three
> > > will likely distort the air bubbles and tissue around them, however.
> > > Plunging works well, as long as the plunge is made very rapidly, and
> > > no time is spent hanging about in the cold -- well below freezing --
> > > nitrogen atmosphere above the cryogen.
> > > Many people plunge into some organic fluid like ethane which is held
> > > in a container in liquid nitrogen. This works, but it has a few
> > > issues: the cryogen is usually warmer than the LN2, so the freezing is
>
> > > not as rapid as it needs to be, and such organic cryogens are
> > > flammable or explosive when they warm up. Especially since, being near
>
> > > LN2 temperatures, they're enriching themselves in oxygen from the air
> > > (liquid air is warmer than liquid nitrogen). They can be handled
> > > safely, but this does require some thought.
> > > A better way is to plunge into slush nitrogen -- LN2 near the freezing
>
> > > point, instead of near the boiling point. This is about 14 deg C
> > > colder than LN2, so freezes samples faster, and so gives a greater
> > > depth of good freezing. It's also nitrogen, so there's no flammable
> > > gas to deal with (nor bureaucrats upset about flammable gases in your
> > > lab). It's easy to produce: just put a beaker full in a small to
> > > medium size vacuum desiccator, and pull a vacuum with a high capacity
> > > pump, like a dual-stage rotary pump. One used to rough out a large EM
> > > column usually works.
> > >
> > > Phil
> > >
> > > We're exploring a treatment in which small short-lived (perhaps on the
>
> > > order of seconds) gas bubbles may be generated in tissue. To this
> > > end, we're interested in learning the 'instantaneous' state of the
> > > tissue immediately after treatment with regard to the presence of any
> > > bubbles. Currently, our experiments utilize rabbit muscle. Are there
> > > any known histological sample preparation/analysis procedures that
> > > have been used to preserve/observe resident gas bubbles? We have
> > > considered a flash freezing followed by staining and/or electron
> > > microscopy. However, our expertise is not in histology, so we'd very
> > > much appreciate any suggestions and/or details regarding possible
> > > sample preparation. Thanks.
> > > Wayne
> > > --
> >
> >
> > /**********************/
> >
> > /Wayne Kreider/
> >
> > /University of Washington/
> >
> > /Applied Physics Lab / CIMU/
> >
> > /106 Old Fisheries/
> >
> > /206-543-1324/
> >
> >
> >
> > ------------------------------
> >
> > Message: 10
> > Date: Tue, 2 Nov 2004 12:55:07 -0500
> > From: tdobersztyn <@t> chmca.org
> > Subject: [Histonet] Re: Polyoma Virus
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <OFD41B23A3.1BE9AB68-ON85256F40.006215F7 <@t> chmca.org>
> > Content-Type: text/plain; charset=US-ASCII
> >
> >
> >
> >
> >
> >
> >
> >
> > Debra-
> > BK, Polyoma virus, can be demonstrated beautifully with Electron
> > Microscopy.
> > That is, if you have access to an EM lab.
> > I am frequently processing renal core biopsies to r/o BK.
> > If you would like assistance-
> > let me know.
> >
> >
> >
> > Date: Mon, 1 Nov 2004 16:16:43 -0500
> > From: Browning Deb <browning <@t> HHSC.CA>
> > Subject: [Histonet] polyoma virus, SV40
> > To: "'histonet <@t> lists.utsouthwestern.edu'"
> > <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID:
> >
> > <3AADFB88753AD31189C100902786B91C0E27851F <@t> hch_nt_exchange.hhsc.ca>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> > Is anyone out there using an antibody against polyoma virus, SV40; BK;
> or
> > JC, against human tissue for demonstrating kidney rejection? If so,
> could
> > you share the details, thanks.
> >
> > Debra Browning, ART
> > Technical Specialist, Immunohistochemistry
> > Anatomic Pathology
> > Hamilton Health Sciences
> > phone: (905) 527-4322 ext 46131
> > e-mail: browning <@t> hhs
> >
> >
> >
> >
> >
> > Theresa R Dobersztyn HT ASCP
> > Electron Microscopy Laboratory
> > Department of Pathology
> >
> > Akron Children's Hospital
> > 1 Perkins Square
> > Akron, Ohio 44308
> >
> > 330-543-8279
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 11
> > Date: Tue, 2 Nov 2004 20:06:49 -0500
> > From: <lpwenk <@t> sbcglobal.net>
> > Subject: Re: [Histonet] AFB Stain
> > To: "Etheridge, Sandra AGF:EX" <Sandra.Etheridge <@t> gems8.gov.bc.ca>,
> > <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <022801c4c141$65f66ca0$6639d445 <@t> domainnotset.invalid>
> > Content-Type: text/plain; charset="iso-8859-1"
> >
> > Check with your microbiology department. They are probably doing it
> > already.
> >
> > One difference I've found between doing this procedure on smears vs.
> > fixed/processed tissue - with smears, there is a 5 minute acid-alcohol
> > decolorization. With fixed/processed/paraffin embedded tissue, reduce
> this
> > to 2-3 seconds. The AFB cells walls have been compromised with the
> > fixative
> > and alcohols and xylene of the processing.
> >
> > Peggy A. Wenk, HTL(ASCP)SLS
> > William Beaumont Hospital
> > Royal Oak, MI 48073
> >
> > ----- Original Message -----
> > From: "Etheridge, Sandra AGF:EX" <Sandra.Etheridge <@t> gems8.gov.bc.ca>
> > To: <histonet <@t> lists.utsouthwestern.edu>
> > Sent: Tuesday, November 02, 2004 12:57 PM
> > Subject: [Histonet] AFB Stain
> >
> >
> > > Hello, all,
> > >
> > > Does anyone have a current method for acid fast bacilli using
> > > auramine-rhodamine? Is this a fluorescent stain?
> > >
> > > Thanks in advance.
> > >
> > > Sandra Etheridge
> > >
> > > BC Ministry of Agriculture, Food & Fisheries
> > > Animal Health Center, Histology
> > > Abbotsford, BC
> > > Canada
> > >
> > > _______________________________________________
> > > Histonet mailing list
> > > Histonet <@t> lists.utsouthwestern.edu
> > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 12
> > Date: Wed, 03 Nov 2004 12:33:41 +0200
> > From: Marshall <marshall <@t> cormack.uct.ac.za>
> > Subject: [Histonet] Merkel cell staining
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <4188B405.A7723577 <@t> cormack.uct.ac.za>
> > Content-Type: text/plain; charset=us-ascii
> >
> > Hi all
> > I am looking to demonstrate Merkel cells in skin. I can not find any
> > reference in my textbooks to their demonstration although I have not
> > used the internet. Also I have tried silver stains for demoing free
> > nerve endings in skin with not much success. The immunocytochemistry for
> > neuro filament protein was a bit better but not great. Is there anyway I
> > can improve on this?
> >
> >
> >
> > ------------------------------
> >
> > Message: 13
> > Date: Wed, 03 Nov 2004 09:41:53 -0500
> > From: "Peter Rippstein" <prippstein <@t> ottawaheart.ca>
> > Subject: [Histonet] LR White vs LR Gold
> > To: <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <s188a7f4.031 <@t> mail.ottawaheart.ca>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Hello Colleagues
> >
> > Have any of you had the opportunity to compare EM immunogold labelling
> > efficiency of LR White vs LR Gold acrylic resins? Protocols would be
> > helpful.
> > Many thanks.
> >
> > Peter
> >
> > Peter Rippstein ART, MLT
> > University of Ottawa Heart Institute
> > 40 Ruskin St., Room H453A
> > Ottawa, Ontario
> > Canada, K1Y 4W7
> >
> > Tel: (613) 761-5282
> > Fax: (613) 761-5281
> > email: prippstein <@t> ottawaheart.ca
> > -------------- next part --------------
> > BEGIN:VCARD
> > VERSION:2.1
> > X-GWTYPE:USER
> > FN:Peter Rippstein
> > EMAIL;WORK;PREF;NGW:PRippstein <@t> ottawaheart.ca
> > N:Rippstein;Peter
> > END:VCARD
> >
> >
> > ------------------------------
> >
> > Message: 14
> > Date: Wed, 3 Nov 2004 08:54:40 -0600
> > From: Sharon Allen <SAllen <@t> exchange.hsc.mb.ca>
> > Subject: [Histonet] RE: Modified Gomori's Trichrome
> > To: 'Bruce Abaloz' <brucea <@t> unimelb.edu.au>, "Histonet (E-mail)"
> > <histonet <@t> pathology.swmed.edu>
> > Message-ID:
> > <304B5A264EC9974E8121B0EA14A9C3936619BA <@t> hsc01mx1.hsc.mb.ca>
> > Content-Type: text/plain; charset="us-ascii"
> >
> > Hi Bruce,
> > I know the Gomori's Trichrome isn't specific for mitochondria but it
> > should
> > stain red if the test is working properly. I have been doing this stain
> > routinely for 4 different Neuropathologists & am now doing the muscles
> for
> > another Neuropathologist who is apparently an expert in the field &
> > extremely fussy. They all are of the same opinion. Having the
> mitochondria
> > stained seems to be imperative for an acceptable stain. Using yet
> another
> > stain on the muscle bx's is not an option. This is why I am attempting
> to
> > find the key to the "perfect Gomori's Trichrome stain". Using the
> > celestine
> > blue before the Mayer's has worked well for staining the mitochondria (I
> > have done 40 of them), but I have no explanation why. My theory is very
> > rusty & hoped someone with more knowledge than I could explain the
> reason
> > to
> > me. I think it may have something to do with Celestine Blue staining DNA
> &
> > being a oxazine dye. Also chromoptrope 2R being a metachromatic stain.
> > I would appreciate any help you can give me.
> > Thanks for your reply,
> > Sharon
> >
> > -----Original Message-----
> >
> > This e-mail and/or any documents in this transmission is intended for
> the
> > address(s) only and may contain legally privileged or confidential
> > information. Any unauthorized use, disclosure, distribution, copying or
> > dissemination is strictly prohibited. If you receive this transmission
> in
> > error, please notify the sender immediately and return the original.
> >
> > ------------------------------
> >
> > Message: 15
> > Date: Wed, 03 Nov 2004 10:32:03 -0500
> > From: "Amy Kozer" <alkozer <@t> piadc.ars.usda.gov>
> > Subject: [Histonet] Envision System with Vector Red Background
> > To: <histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <s188b672.089 <@t> FSNAA-2.naa.ars.usda.gov>
> > Content-Type: text/plain; charset=US-ASCII
> >
> > I've tried using Vector Red with Envision system for permanance and have
> > encountered a lot of background and precipitate.
> > I've been staining frozen sections of epithelium fixed with acetone.
> > Has anyone had any experience with this? Remedies?
> >
> > Thank you
> > Amy Kozer
> > PIADC, ARS, USDA
> >
> >
> >
> > ------------------------------
> >
> > Message: 16
> > Date: Wed, 03 Nov 2004 10:52:46 -0500
> > From: "Richard Cartun" <Rcartun <@t> harthosp.org>
> > Subject: Re: [Histonet] Merkel cell staining
> > To: <marshall <@t> cormack.uct.ac.za>,<histonet <@t> lists.utsouthwestern.edu>
> > Message-ID: <s188b88d.075 <@t> harthosp.org>
> > Content-Type: text/plain; charset=US-ASCII
> >
> > Try IHC for cytokeratin 20.
> >
> > 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
> >
> > >>> Marshall <marshall <@t> cormack.uct.ac.za> 11/03/04 05:33AM >>>
> > Hi all
> > I am looking to demonstrate Merkel cells in skin. I can not find any
> > reference in my textbooks to their demonstration although I have not
> > used the internet. Also I have tried silver stains for demoing free
> > nerve endings in skin with not much success. The immunocytochemistry
> > for
> > neuro filament protein was a bit better but not great. Is there anyway
> > I
> > can improve on this?
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> >
> >
> > ------------------------------
> >
> > Message: 17
> > Date: Wed, 03 Nov 2004 11:09:31 -0500
> > From: Rita Angel <RITA.ANGEL <@t> UC.EDU>
> > Subject: [Histonet] B & D steamer
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <5.1.0.14.2.20041103110800.00b24e08 <@t> ucmail3.uc.edu>
> > Content-Type: text/plain; charset="us-ascii"; format=flowed
> >
> > Thanks to everyone for their input on the type of containers you use in
> > the
> > steamer. I'll try the plastic coplin jars!!
> >
> > Thanks again,
> >
> > Rita Angel HT (ASCP)
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 18
> > Date: Wed, 3 Nov 2004 08:52:46 -0800 (PST)
> > From: Histology SLU <sluhisto <@t> yahoo.com>
> > Subject: [Histonet] Polyoma BK virus controls
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <20041103165246.4952.qmail <@t> web51001.mail.yahoo.com>
> > Content-Type: text/plain; charset=us-ascii
> >
> > Does anyone have BK virus controls that you would be willing to trade
> for
> > something we may have that you would need? Any help in locating
> control
> > blocks or slides would be greatly appreciated. Thanks.
> >
> > Susan
> >
> >
> > ---------------------------------
> > Do you Yahoo!?
> > Check out the new Yahoo! Front Page. www.yahoo.com/a
> >
> > ------------------------------
> >
> > Message: 19
> > Date: Wed, 3 Nov 2004 11:46:59 -0600
> > From: "Grant, Debra" <DRG <@t> Stowers-Institute.org>
> > Subject: [Histonet] BMP7
> > To: <histonet <@t> pathology.swmed.edu>
> > Message-ID: <1112618780-192831059 <@t> pathology.swmed.edu>
> > Content-Type: text/plain; charset="us-ascii"
> >
> >
> > Has anyone used BMP7 antibody on mouse? If so, could you please send me
> > all the information, company, catalog #, protocol etc.
> >
> > Thanks in advance!
> >
> > Debby Grant
> > Research Technician II
> > Histology Core Facility
> > Stowers Institute for Medical Research
> > 1000 E. 50th Street
> > Kansas City, MO 64110
> > drg <@t> stowers-institute.org
> >
> >
> >
> >
> > ------------------------------
> >
> > Message: 20
> > Date: Wed, 03 Nov 2004 09:52:01 -0800
> > From: "Ant S." <neuroant <@t> hotmail.com>
> > Subject: [Histonet] Thanks for the Teasing
> > To: histonet <@t> lists.utsouthwestern.edu
> > Message-ID: <BAY15-F22ikaTAbNF440001aced <@t> hotmail.com>
> > Content-Type: text/plain
> >
> >
> > Thanks for the great tips on Nerve Teasing. I have quite a few
> good
> > ideas to experiment with now. I appreciate the help. This is a
> very
> > informative community (and mostly cheerful, too!)
> >
> > Thanks,
> >
> > Antoinette Swensson
> > Univeristy of Washington/Harborview Medical Center
> > Neuropathology
> > _________________________________________________________________
> >
> > [1]Find the music you love on MSN Music. Start downloading now!
> >
> > References
> >
> > 1. http://g.msn.com/8HMBENUS/2731??PS=47575
> >
> >
> > ------------------------------
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >
> > End of Histonet Digest, Vol 12, Issue 4
> > ***************************************
> >
> >
>
>
>
> ------------------------------
>
> Message: 2
> Date: Wed, 3 Nov 2004 13:04:05 -0700
> From: "Elizabeth Chlipala" <lizchlipala <@t> premierhistology.com>
> Subject: [Histonet] TB block
> To: <histonet <@t> pathology.swmed.edu>
> Message-ID: <000501c4c1e0$45969cb0$76d48a80 <@t> AMY>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello everyone
>
> I was wondering if there is anyone out there that could spare a block
> that contains mycobacterium tuberculosis, and be willing to send that to
> me. I would be willing to pay for the cost of shipping. I'm currently
> trying to characterize an antibody to TB and the control block of
> mycobacterium avium that I have will not work for my application.
>
> Thanks
>
> Liz
>
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Premier Laboratory, LLC
> P.O. Box 18592
> Boulder, Colorado 80308
> Office: (303) 735-5001
> Fax: (303) 735-3540
> liz <@t> premierlab.com
> www.premierlab.com
>
> Ship to Address:
> Premier Laboratory
> University of Colorado
> MCDB, Room A3B40
> Boulder, Colorado 80309
>
>
>
> ------------------------------
>
> Message: 3
> Date: Wed, 3 Nov 2004 19:45:19 -0300
> From: "Hernan Aldana" <haldana <@t> unimoron.edu.ar>
> Subject: [Histonet] Fixation for carbohydrates
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000001c4c1f6$cc542a40$389559c8 <@t> b3w6zzmtb6juvbs>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hello all
> I need the best fixation method for preserve hyaluronic acid in paraffin
> sections. Formaldehyde, Bouin, Zencker, B5? Thanks in advance
>
>
> Dr. Hernán J. Aldana Marcos
> Facultad de Medicina. Universidad de Morón
> Machado 914. B1708JPD. Buenos Aires. Argentina
> e-mail alternativo hernanjavier <@t> yahoo.com
> web: http://hjaldanamarcos.bravepages.com
> http://www.ht.org.ar
>
>
>
>
>
>
> ------------------------------
>
> Message: 4
> Date: Wed, 3 Nov 2004 18:43:22 EST
> From: JCarpenter764 <@t> aol.com
> Subject: [Histonet] practical exam...
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <ff.59e466d.2ebac71a <@t> aol.com>
> Content-Type: text/plain; charset="US-ASCII"
>
> has anyone heard about the practical and wether they passed?
>
>
> ------------------------------
>
> Message: 5
> Date: Wed, 3 Nov 2004 17:49:34 -0600
> From: "maryjohnson" <beingmary53 <@t> ev1.net>
> Subject: [Histonet] exam
> To: <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <003501c4c1ff$c64cf510$b248bbd0 <@t> ugly>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi all I to am wating to hear about the practial exam.
>
>
> ------------------------------
>
> Message: 6
> Date: Wed, 03 Nov 2004 18:51:51 -0500
> From: Amos Brooks <amosbrooks <@t> earthlink.net>
> Subject: [Histonet] Using expired reagents in IHC
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <6.0.0.22.0.20041103182925.01afbe90 <@t> pop.earthlink.net>
> Content-Type: text/plain; charset="us-ascii"; format=flowed
>
> Hi,
> We always get new antibodies before the old expire. We only have
> a
> few expired ones kicking around. The Big High Muckety-Mucks (somewhat
> understandably) have determined that the cost of the fines and dealing
> with
> battling the inspectors would be worse, in the short term, than the cost
> of
> purchasing just enough of the antibodies and not dealing with the extra
> tech time involved in testing the same antibody monthly.
> I can see the logic of either side of the coin. It may be easy to
>
> overlook a reagent quality gradually decreasing. Repeated testing of the
> reagent costs time & money too. And the risk of the fines looming over
> your
> head is enough to make one hesitate. On the other hand the money saved on
> keeping antibodies around for decades is appealing. I guess you could say
> we're erring on the side of caution ... bloody expensive caution.
> On another note, has anyone noticed that the listed shelf life of
>
> these reagents is shrinking lately. I just got one in today that has about
>
> 9 months until it expires. It is enough to make one wonder if the antibody
>
> companies are pushing for these regulations. I mean, really, where do they
>
> come up with these dates anyway?
> Have a great day,
> Amos Brooks
>
>
>
> At 10:54 AM 11/2/2004, you wrote:
> >Message: 3
> >Date: Mon, 01 Nov 2004 14:26:32 -0500
> >From: "Richard Cartun" <Rcartun <@t> harthosp.org>
> >Subject: [Histonet] Using expired reagents in IHC
> >To: <histonet <@t> lists.utsouthwestern.edu>
> >Message-ID: <s18647a7.003 <@t> harthosp.org>
> >Content-Type: text/plain; charset=US-ASCII
> >
> >It has been brought to my attention that the CAP Laboratory checklist
> >has been revised as of 9/30/04 and now includes a question, "Are all
> >immunohistochemical reagents used within their indicated expiration
> >dates?". I have always believed that it is acceptable to use expired
> >regents (mainly primary antibodies) as long as their reactivity is
> >acceptable and documented. Obviously, the expiration date that
> >manufactures use is not an accurate reflection of the reagent's
> >likelihood of poor performance. Does anyone have information on this
> >change? Thank you.
> >
> >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
>
>
>
>
>
> ------------------------------
>
> Message: 7
> Date: Wed, 3 Nov 2004 16:49:22 -0800 (PST)
> From: Gareth Davis <mrsgbd2001 <@t> yahoo.com>
> Subject: [Histonet] Re: Exam
> To: Histonet <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <20041104004922.8210.qmail <@t> web52703.mail.yahoo.com>
> Content-Type: text/plain; charset=us-ascii
>
> >From what I've heard, the results for the pratical exam will not be out
> until late November or early December.
> Gareth
>
>
> ---------------------------------
> Do you Yahoo!?
> Check out the new Yahoo! Front Page. www.yahoo.com/a
>
> ------------------------------
>
> Message: 8
> Date: Wed, 3 Nov 2004 20:30:29 -0500
> From: "Denise Bland-Piontek" <dbpiontek <@t> clinomicsbio.com>
> Subject: [Histonet] test
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <145F33B84793CF488578F86355B32E6912FBA4 <@t> mail.clinomicsbio.com>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
>
>
> ------------------------------
>
> Message: 9
> Date: Wed, 3 Nov 2004 20:29:39 EST
> From: Dndsomi <@t> aol.com
> Subject: Re: [Histonet] Re: Exam
> To: mrsgbd2001 <@t> yahoo.com, Histonet <@t> lists.utsouthwestern.edu
> Message-ID: <1a7.2a74b0d2.2ebae003 <@t> aol.com>
> Content-Type: text/plain; charset="US-ASCII"
>
> I sent my practical in Sept. 2003 and got my results Jan. 15, 2004.
>
>
> ------------------------------
>
> Message: 10
> Date: Wed, 3 Nov 2004 21:54:23 EST
> From: CrochiereSteve <@t> aol.com
> Subject: [Histonet] EGFR
> To: PatPatterson <@t> mhd.com, histonet <@t> pathology.swmed.edu
> Message-ID: <1d8.2f65199e.2ebaf3df <@t> aol.com>
> Content-Type: text/plain; charset="US-ASCII"
>
> Pat,
> USLabs in California
>
> Steven M. Crochiere, HT(ASCP)
> Histology Supervisor
> LifePath Partners @ Mercy Medical Center
> Springfield, MA 01104
>
>
> ------------------------------
>
> Message: 11
> Date: Thu, 04 Nov 2004 13:48:12 +0000
> From: Chris.Goodall <@t> bristol.ac.uk
> Subject: [Histonet] Timm silver sulphide method for light microscopic
> localisation of heavy metals.
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <1099576092.418a331c8b602 <@t> webmail.bris.ac.uk>
> Content-Type: text/plain
>
>
> Dear All,
> I have been asked to try the Timm method on some FFPE samples of
> human brain. The papers I have read give methods for perfusing rat
> brain with sodium sulphide solution followed by 3% gluteraldehyde in
> 0.15M phosphate buffer, and further treatment with sodium sulphide
> solution, or, post mortem brain samples snap frozen and frozen
> sectioned. However health and safety rules here do not permit frozen
> sectioning of unfixed human brain, or the use of gluteradehyde in the
> embalming room, so I am going to attempt this method on formalin fixed
> samples.My question is, has anyone experience of this method or any
> suggestions? It is mentioned that autopsy material left in situ for one
> or two days post mortem with no fixation may have enough endogenous
> sulphide ions or sulphide groups in the tissue due to autolytic
> activity and sulphide treatment may not be necessary, or if it is
> necessary, does anyone have experience of timing in the sulphide
> solution and is post fixation necessary after the initial formalin
> fixation. The paper also mentions that to prevent loss of
> metallosulphides present in the tissue the temperature of the wax
> should not exceed 50C which means the use of the dreaded low
> temperature wax, has anyone used conventional paraffin wax and got away
> with it?
> Sorry for the long request,and thank you,
> Chris Goodall
>
>
>
>
> ------------------------------
>
> Message: 12
> Date: Thu, 4 Nov 2004 10:03:38 -0600
> From: ajennings <@t> unmc.edu
> Subject: [Histonet] Histonet 2005 Buttons
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> <OF04E01F03.C7F5D4D1-ON86256F42.0057AA6D-86256F42.005838FE <@t> unmc.edu>
> Content-Type: text/plain; charset=US-ASCII
>
>
>
>
>
> This is it...... I am going to let people continue to submit their designs
> until next Friday and then I will be presenting the designs to our working
> group to help me decide which one we will use for the Official 2005
> Histonet button. If you have been saving your stuff for last minute.....we
> are in the 60 second count down for submissions.
>
> Please include a space for writing a name, the URL and a place for the
> company who sponsors the buttons.
>
> Thanks a bunch for all the ones we have received it is fun to see the
> talent and the involvement of the community
> Anita
>
>
>
>
>
> ------------------------------
>
> Message: 13
> Date: Thu, 4 Nov 2004 16:16:08 GMT
> From: "mprice26 <@t> juno.com" <mprice26 <@t> juno.com>
> Subject: [Histonet] RE: Job Openings- Texarkana, TX
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <20041104.081617.26898.15797 <@t> webmail28.nyc.untd.com>
> Content-Type: text/plain
>
>
> Looking for HT (ASCP) Registered Histotech at Pathology Services of
> Texarkana. Great Location to raise a family. Day shift.
>
> Also looking for a temporary histotech for a couple of months.
>
> Contact Marsha Price @ mprice26 <@t> juno.com.
>
> Marsha
>
> ________________________________________________________________
> Juno Platinum $9.95. Juno SpeedBand $14.95.
> Sign up for Juno Today at http://www.juno.com!
> Look for special offers at Best Buy stores.
>
>
>
> ------------------------------
>
> Message: 14
> Date: Thu, 04 Nov 2004 11:21:19 -0500
> From: John Kiernan <jkiernan <@t> uwo.ca>
> Subject: Re: [Histonet] Timm silver sulphide method for light
> microscopiclocalisation of heavy metals.
> To: Chris.Goodall <@t> bristol.ac.uk
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <418A56FF.69DDC8C6 <@t> uwo.ca>
> Content-Type: text/plain; charset=us-ascii
>
> The perfusion of sodium sulphide is necessary
> to precipitate metal ions (principally zinc),
> which are otherwise soluble. Relying on autolysis
> to generate sulphide and precipitate the zinc
> doesn't make sense because the zinc ions would
> have diffused away from their original synaptic
> sites, and the tissue would be decomposing
> anyway. Timm's method simply won't work (or not
> in any meaningful way) on formaldehyde-fixed
> paraffin-embedded specimens.
>
> Glutaraldehyde is not the only permissible
> fixative after perfusing the buffered Na2S.
> An alternative is to use an alcoholic fixative
> that has been saturated with H2S gas. It's
> unlikely that your health & safety regulations
> will allow this if they won't let you take a
> little jar of glutaraldehyde into the PM room!
>
> The best account of modern Timm staining is
> probably Danscher,G & Zimmer,J (1978)
> Histochemistry 55: 27-40. Danscher has also
> published many more recent papers on
> histochemical detection of zinc, mercury and
> other metals with sulphide-silver methods.
>
> John Kiernan
> London, Canada.
> ___________________________
> Chris.Goodall <@t> bristol.ac.uk wrote:
> >
> > Dear All,
> > I have been asked to try the Timm method on some FFPE samples of
> > human brain. The papers I have read give methods for perfusing rat
> > brain with sodium sulphide solution followed by 3% gluteraldehyde in
> > 0.15M phosphate buffer, and further treatment with sodium sulphide
> > solution, or, post mortem brain samples snap frozen and frozen
> > sectioned. However health and safety rules here do not permit frozen
> > sectioning of unfixed human brain, or the use of gluteradehyde in the
> > embalming room, so I am going to attempt this method on formalin fixed
> > samples.My question is, has anyone experience of this method or any
> > suggestions? It is mentioned that autopsy material left in situ for one
> > or two days post mortem with no fixation may have enough endogenous
> > sulphide ions or sulphide groups in the tissue due to autolytic
> > activity and sulphide treatment may not be necessary, or if it is
> > necessary, does anyone have experience of timing in the sulphide
> > solution and is post fixation necessary after the initial formalin
> > fixation. The paper also mentions that to prevent loss of
> > metallosulphides present in the tissue the temperature of the wax
> > should not exceed 50C which means the use of the dreaded low
> > temperature wax, has anyone used conventional paraffin wax and got away
> > with it?
> > Sorry for the long request,and thank you,
> > Chris Goodall
> >
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 15
> Date: Thu, 4 Nov 2004 10:20:44 -0700
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: RE: [Histonet] exam
> To: "'maryjohnson'" <beingmary53 <@t> ev1.net>,
> <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000f01c4c292$a0e4f5f0$83020a0a <@t> IHCTech>
> Content-Type: text/plain; charset="us-ascii"
>
> We just did the grading of over 1100 slide sets for the HT exam last
> weekend in Chicago, I am sure it will take some extra time this go round
> to process all those.
> Patsy
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
> maryjohnson
> Sent: Wednesday, November 03, 2004 4:50 PM
> To: Histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] exam
>
>
> Hi all I to am wating to hear about the practial exam.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 16
> Date: Thu, 4 Nov 2004 10:32:45 -0700
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: RE: [Histonet] BMP7
> To: "'Grant, Debra'" <DRG <@t> Stowers-Institute.org>,
> <histonet <@t> pathology.swmed.edu>
> Message-ID: <001101c4c294$4ba9e3f0$83020a0a <@t> IHCTech>
> Content-Type: text/plain; charset="us-ascii"
>
> Debra,
> I have used Santa Cruz goat BMP-7 (L-19) sc-9305 on a lot of rat and
> human samples but have not tried it on ms. SC lists it as working for
> rat and h, since it is a goat poly it would be easy to try it on your
> ms, ask SC if anyone has reported it as working on ms although I don't
> usually find SC being very helpful for anything not listed being tried.
> For rats I use it at 1:200 for 1 hr. using LSAB+ detection (I wish
> someone would come up with a labeled polymer that works with goat
> antibodies) and pepsin digestion (premade from Phoenix) at 37dc for two
> changes 5 min. ea.
> Patsy
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Grant,
> Debra
> Sent: Wednesday, November 03, 2004 10:47 AM
> To: histonet <@t> pathology.swmed.edu
> Subject: [Histonet] BMP7
>
>
>
> Has anyone used BMP7 antibody on mouse? If so, could you please send me
> all the information, company, catalog #, protocol etc.
>
> Thanks in advance!
>
> Debby Grant
> Research Technician II
> Histology Core Facility
> Stowers Institute for Medical Research
> 1000 E. 50th Street
> Kansas City, MO 64110
> drg <@t> stowers-institute.org
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 17
> Date: Thu, 4 Nov 2004 10:48:19 -0700
> From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
> Subject: [Histonet] bone marrow mush
> To: <Histonet <@t> lists.utsouthwestern.edu>
> Cc: ihcrg <@t> neo.agsci.colostate.edu
> Message-ID: <001701c4c296$79712e40$83020a0a <@t> IHCTech>
> Content-Type: text/plain; charset="us-ascii"
>
> Folks,
> I have a question for you. One of my investigators harvested bone
> marrow by just scrapping it out of the bone cavity, it is not in
> anything (media, anticoagulant, fixative, etc.) they just stored it at
> 4dc as a mush in a tube. They want me now to prepare it so that they
> can see what cells they have. They also want to know if there are any
> stem cells and what the viability is (by viability I mean they are
> wondering if they could use it to grow some of the cells in culture). I
> really don't think I can tell them if they can grow the cells or not.
> Anyway I was thinking of trying to make a cell block from this material
> and process into paraffin or maybe even do frozen sections on the
> pellet. How would you go about making a pellet out of that stuff?
> Would you put some buffer or media or fixative in the tube and spin it
> down and then put it in HISTOGEL or wrap in tissue paper and then
> process?
> Patsy Ruegg
>
>
> ------------------------------
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> End of Histonet Digest, Vol 12, Issue 5
> ***************************************
>
>
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