Re:Brain IHC

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From:Louise Burrell <lburrell@pathbox.wustl.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
Reply-To:
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Dear All:  My extensive experienc with human brain taught me that the
lowest applicable titer and O/N inc ubation with primary works very well
on parqafiin/or frozens of brain

Thanks,
Louise


On Fri, 18 Dec 1998, HistoNet Server wrote:

>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 07:16:06 -0600
> From: Elizabeth Morehead <DREWSB@smtpgw2.musc.edu>
> Subject: DNA/RNA/CYTOPLASM -Reply
>
> test
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 09:00:16 -0600
> From: "Barry, Lilith" <Lilith.Barry@nrc.ca>
> Subject: Re: [IHC on frozen sections of brain]
>
> I use Vector ABC kit routinely on perfused brains and it works very well.
> The few times I have tried to use it on fresh-frozen sections, the
> background was very high. Does anybody know why?
>  Lilith
>
>  ----------
> From: Carol Burden
> To: HistoNet@Pathology.swmed.edu
> Subject: Re: [IHC on frozen sections of brain]
> Date: Wednesday, December 16, 1998 6:26PM
>
> HistoNet@Pathology.swmed.edu wrote:
>
> Hello Everyone,
> Does any of you have a tried and proven protocol for IHC on frozen brain
> sections using preferably the vector ABC-HRP system or any ABC staining
> system?
>
> your prompt reply will be greatly appreciated.
>
> Thanking you in advance.
> Atoska S.Gentry,B.S, HT(ASCP)
>
>
> I'd like this info as well,could you please forward your findings.
> Sincerely Thanks,Carol at Emory
>
>
> ____________________________________________________________________
> More than just email--Get your FREE Netscape WebMail account today at
> http://home.netscape.com/netcenter/mail
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 09:00:40 -0600
> From: vandeplas@aurion.nl (Peter van de Plas)
> Subject: Re: autofluorescence
>
> Ethan Hofmann wrote:
> >I don't know if anybody's seen this yet, but I'm looking for a way to
> >reduce or quench endogenous autofluorescence in murine GALT (gut associated
> >lymphoid tissue).  Are there any successful techniques for this out there?>
> - -----------------------------
> Dear Ethan,
> On aldehyde fixed specimens i always use 0.1% sodium borohydrid in PBS for
> 15 minutes. This treatment not only quenches free aldehyde groups but also
> autofluorescence.
> Kind regards, Peter
>
> ========================================
> Peter van de Plas
> AURION
> Costerweg 5
> 6702 AA Wageningen
> The Netherlands
> phone: (31)-317-497676
> fax: (31)-317-415955
> http://www.aurion.nl
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 09:20:26 -0600
> From: rschoonh@sph.unc.edu
> Subject: Re: Fixative
>
> Mary Kay,
>
> I am assuming that you are reffering to smears?  For that we use cold
> methanol (100%).
>
> - -- Begin original message --
>
> > To All;
> >   Do you think the best fixative for immunos is Acetone no matter what
> > antibody one is using or is their a beter one. Does it definitely make
> > a difference as to the antibody?
> >    Thanks for your input.
> >         Mary Kay Olson
> >         CBN and Anatomy
> >         Loyola University Med. Center
> >         molson2@luc.edu
> >
> >
>
> - -- End original message --
>
> regards,
> Bob
> Robert Schoonhoven
> Laboratory of Molecular Carcinogenesis and Mutagenesis
> Dept. of Environmental Sciences and Engineering
> University of North Carolina
> CB#7400
> Chapel Hill, NC 27599
> Phone
> office 919-966-6343
>    Lab 919-966-6140
>    Fax 919-966-6123
>
> **I'm willing to make the mistakes if someone else is willing to learn
> from them**
>
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 10:15:28 -0600
> From: "Mary L. Verlinde" <verlinde@ahdlms.cvm.msu.edu>
> Subject: ANTIGEN RETRIEVAL
>
>
> I am looking for a recipe for an antigen retrieval acid or buffer for
> paraffin slides to do IHC. Thanks in advance.
>
>
>
> Mary Verlinde
> Animal Health Diagnostic Lab
> MSU
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 10:15:52 -0600
> From: mcauliff@UMDNJ.EDU
> Subject: Re: [IHC on frozen sections of brain]
>
> Carol Burden wrote:
> >
> > HistoNet@Pathology.swmed.edu wrote:
> >
> > Hello Everyone,
> > Does any of you have a tried and proven protocol for IHC on frozen brain
> sections using preferably the vector ABC-HRP system or any ABC staining
> system?
> >
> > your prompt reply will be greatly appreciated.
> >
> > Thanking you in advance.
> > Atoska S.Gentry,B.S, HT(ASCP)
> >
> > I'd like this info as well,could you please forward your findings.
> > Sincerely Thanks,Carol at Emory
> >
> > ____________________________________________________________________
> > More than just email--Get your FREE Netscape WebMail account today at
> http://home.netscape.com/netcenter/mail
>
>
> 	Do you have a specific question or is the whole proceedure in doubt? I
> follow a fairly routine proceedure paying attention to Vector's
> instructions for their reagents. E-mail me off list (unless I hear a
> great clamor for my 'widsom').
>
> Geoff
> - --
> ***************************************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane  Piscataway, NJ 08854
> voice: (732)-235-4583; fax -4029   e-mail: mcauliff@umdnj.edu
> ***************************************************************
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 10:38:36 -0600
> From: mcauliff@UMDNJ.EDU
> Subject: Re: [IHC on frozen sections of brain]
>
> Barry, Lilith wrote:
> >
> > I use Vector ABC kit routinely on perfused brains and it works very well.
> > The few times I have tried to use it on fresh-frozen sections, the
> > background was very high. Does anybody know why?
> >  Lilith
> >
>
> 	I am also very pleased with the Vector kit(s). In perfused tissue
> plasma (and other) proteins are washed out of the tissues while in fresh
> frozen they remain, thus the higher background (I think).
>
> Geoff
> - --
> ***************************************************************
> Geoff McAuliffe, Ph.D.
> Neuroscience and Cell Biology
> Robert Wood Johnson Medical School
> 675 Hoes Lane  Piscataway, NJ 08854
> voice: (732)-235-4583; fax -4029   e-mail: mcauliff@umdnj.edu
> ***************************************************************
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 12:15:44 -0600
> From: "Sebree Linda A." <la.sebree@hosp.wisc.edu>
> Subject: RE: ANTIGEN RETRIEVAL
>
> Mary,
>
> Try 1mM EDTA, pH 8 for anywhere from 5-40" in some type of heating
> apparatus, i.e. water bath, microwave oven, hot plate, pressure cooker, etc.
> We bring our slides and buffer up to just boiling, then incubate them in a
> hot (91-94 degrees C) water bath for the duration.  We used to cool under
> running tap water for 20" before staining but recently we've tried going
> right from retrieval to staining with no discernible differences; sure saves
> time!
>
> Linda
>
> Linda Sebree, HT
> University of Wisconsin Hospital & Clinics
> Department of Laboratory Medicine
> IHC/ISH Laboratory
> A4/204-2472
> 600 Highland Ave.
> Madison, WI  53792-2472
>
> Phone:  (608)265-6596
> FAX:  (608)263-1568
>
> > -----Original Message-----
> > From:	Mary L. Verlinde [SMTP:verlinde@ahdlms.cvm.msu.edu]
> > Sent:	Thursday, December 17, 1998 1:07 PM
> > To:	'HISTONET@PATHOLOGY.SWMED.EDU'
> > Subject:	ANTIGEN RETRIEVAL
> >
> >
> > I am looking for a recipe for an antigen retrieval acid or buffer for
> > paraffin slides to do IHC. Thanks in advance.
> >
> >
> >
> > Mary Verlinde
> > Animal Health Diagnostic Lab
> > MSU
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 12:46:23 -0600
> From: bkrummre@ingham.k12.mi.us
> Subject: unscubscribe
>
>
>
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 14:30:40 -0600
> From: "Plummer, Timothy B." <Plummer.Timothy@mayo.edu>
> Subject: Griffonia Simplicifolia (GSA IB4) Lectin
>
> Hello,
>
> Does anyone have a good protocol for using GSA IB4-FITC in paraffin sections
> (formalin fixed).  Thanks in advance.
>
> Tim
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 15:00:43 -0600
> From: "Fran Adams" <franadams7@hotmail.com>
> Subject: Please unsubscribe
>
> Please unsubscribe.
>
> Thank you!  Happy Holidays!
>
> ______________________________________________________
> Get Your Private, Free Email at http://www.hotmail.com
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 16:15:52 -0600
> From: "bbroders@unlnotes.unl.edu" <bbroders@unlnotes.unl.edu>
> Subject: FW: Slide costs
>
> Thought I'd forward this to this list.  Seems to me I saw someone else ask
> a similar question here on histonet, but I didn't pay much attention to the
> replies.
> Thanks,
> Bruce
>
> - -----Original Message-----
> From:	Dr. Bruce Akey [SMTP:bakey@vdacs.state.va.us]
> Sent:	Thursday, December 17, 1998 4:00 PM
> To:	American Assoc of Vet Lab Diagnosticians
> Subject:	Slide costs
>
>
>
>
> Has anyone gone through the tedious process of figuring what it costs/slide
> to produce an H&E slide?  Just the materials/reagents costs is primarily
> what I need, without the manpower or instrument depreciation etc...  If you
> have done this within the last year and would be good enough to share that
> information I would greatly appreciate it.
>
> TIA
>
> Bruce
>
> ~~~~~~~~~~~~~~~~~~~~~~~
> Bruce L. Akey, MS, DVM
> Chief, Office of Laboratory Services
> Virginia Dept. Agriculture and Consumer Services
> (804) 786-9202
> bakey@vdacs.state.va.us
>
>
> ----------------------------------------------------------------------
>
> Date: 17 Dec 1998 22:30:18 -0600
> From: Katri Tuomala <katri@istar.ca>
> Subject: Re: ANTIGEN RETRIEVAL
>
> Mary L. Verlinde wrote:
> >
> > I am looking for a recipe for an antigen retrieval acid or buffer for
> > paraffin slides to do IHC. Thanks in advance.
> >
> > Mary Verlinde
> > Animal Health Diagnostic Lab
> > MSU
> Hi Mary,
> Citrate buffer pH 6.0 is commonly used in microwave retrieval of
> formalin fixed, paraffin embedded material. I make my own by dissolving
> 2.1 g of citric acid in 1000 ml dist.water and bringing the pH up to 6.0
> by adding 2M sodium hydroxide. I use microwavable pressure cooker, which
> allows continued heating without excessive evaporation of the buffer.
> Bring the slides in buffer to boiling 10-15 minutes on HIGH. Immediately
> add another 2-3 minutes on HIGH. Then let slides sit in hot solution for
> 20 minutes. Continue with your method. You can do this in coplin jars,
> if you don't have a pressure cooker, but you have to add more buffer
> every 5 minute intervals of heating to avoid the slides from drying out.
> To obtain reproducible results try to keep the amount of the solution
> and number of slides constant. Hope this is helpful, good luck! Katri.
>
> Katri Tuomala
> Department of Pathology
> St.Joseph's Hospital
> Hamilton, Ontario, Canada
>
>
> Here are the messages received yesterday!
>
>




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