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-----Original Message-----
From: HistoNet Server <HistoNet@Pathology.swmed.edu>
To: HistoNet Server <HistoNet@Pathology.swmed.edu>
Date: Thursday, December 31, 1998 1:08 AM
Subject: Daily Digest


>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 00:07:24 -0600
>From: "Patricia M. Karlisch" <pkarlisch@psghs.edu>
>Subject: Daily Digest -Reply
>
>I will be out of the office from Dec 28th - Jan3.  Please leave a message
and
>I will return your e-mail as soon as possible. If it is urgent I can be
>reached at 271-0665.  Happy Holidays.  Pat
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 09:46:05 -0600
>From: Elizabeth Hayes <ehayes@bcm.tmc.edu>
>Subject: unsubscibe
>
>unsubscribe
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 10:31:27 -0600
>From: lpwenk@mail.netquest.com
>Subject: Re: Cathy Sanderson-Mayton
>
>andrea_kelly@CCGATEWAY.AMC.EDU wrote:
>>
>>      I'm looking for Cathy's new email address.Thank you!
>
>Found her on the Histotech's Home Page "Directory"
>
> http://www.histology.to
>
>Go to Directory. Go to first alphabet letter of last name.
>Start scrolling.
>
>There is also a submission page, if you would like your
>email address listed. (and your snail mail address, if you
>like).
>
>P.S. Found Cathy under Mayton-Sanderson, not Sanderson-Mayton.
>- --
>Peggy A. Wenk, HTL (ASCP)
>Anatomic Pathology
>Wm. Beaumont Hospital
>3601 W. 13 Mile Rd.
>Royal Oak, MI 48073-6769
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 11:01:11 -0600
>From: bresee@pilot.msu.edu
>Subject: Re: Myeloperoxidase Staining
>
>To: Clif Chapman, Cathy Fragiskatos, and any other interested parties.
>
>What kind of antibody do you use against myeloperoxidase?  Does it stain
>neutrophils?
>
>I am trying the myeloperoxidase assay from the AFIP manual with little
>success.  Here's what I'm trying to do:
>
>I am trying to do a stain that will allow easy detection for
>quantitation of all the neutrophils in formalin-fixed, paraffin-embedded
>tissues so I can tell them apart from eosinophils and not rely solely on
>the morphology of the nucleus.  We have found that a May-Grunwald stain
>is great for the eos.  We probably could count the neutrophils in these
>May-Grunwald stained sections, but I would like to find something that
>makes those neutrophils really stand out.  I am not planning on doing
>both stains on one slide, but on serial sections.  However, a good  IHC
>stain for neutrophils with a May-Grunwald counterstain would be awesome!
>
>Thanks in advance for any info you might have!
>
>
>Catherine "Katie" Bresee Bennett
>218G Food Safety Toxicology Building
>Department of Pathology
>Michigan State University
>East Lansing, MI 48824
>
>ph:  (517) 432-4940
>fx:   (517) 353-9902
>
>bresee@pilot.msu.edu
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 11:45:50 -0600
>From: "Hagerty, Marjorie A." <mhagerty@emc.org>
>Subject: RE: Strange new artifact
>
>Hi Lynn and fellow histonetters,
>
>Thanks for your reply. I'm pretty sure it is not formalin pigment, it
>looks nothing like formalin pigment to me. After reading your post it
>occurred to me that I should recut the block and see if it is still
>there or if it happened after processing. Would you be willing to look
>at the slide? I would be very appreciative if you would review a slide.
>Is there anyone else out there in cyberspace who wants to take a shot at
>this. I would be happy to send anyone who would like to participate a
>slide with my "so-called" artifact on it. We could then compare notes.
>Any help would be appreciated and as you stated Lynn, it would be hard
>to make a guess without actually looking at the slide. Any takers please
>send me your address and I'll send it right out to you!
>
>Thanks.
>
>Marg
>EMC, Rancho Mirage, CA
>> -----Original Message-----
>> From: Lynn Gardner [SMTP:gardnerl@horus.ophth.uiowa.edu]
>> Sent: Wednesday, December 30, 1998 4:56 AM
>> To: Hagerty, Marjorie A.
>> Subject: Re: Strange new artifact
>>
>> Could it possibly be formalin pigment? How long are the specimens in
>> fixative before decalcification? Have you checked the date on your
>> decal
>> solution to assure that it is still good? These are possibilities but
>> would
>> have to see the pigmentation to really know what it is.
>>
>> Good Luck!
>>
>> At 01:55 PM 12/28/98 -0800, you wrote:
>> >Hi Everyone,
>> >
>> >We process a lot of decalcified tissue, hips, knees, etc. A new
>> artifact
>> >(new to us at least) has recently appeared. It is a brownish
>> >pigmentation in the cartilage and seen microscopically. Has anyone
>> ever
>> >seen this before? Does anyone have a clue as to what may be causing
>> it?
>> >
>> >Nothing has changed with the way we decalcify. We use formical 4 as a
>> >decalcifier. A general procedure that varies somewhat with individual
>> >specimens is to: put in formalin for 3 days, cut with saw and put
>> back
>> >in formalin for 2 days, then put in formical 4 for 3-4 days.
>> >
>> >Hope someones knows!
>> >
>> >Thanks,
>> >Marg
>> >EMC, Rancho Mirage, CA
>> >
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 15:09:24 -0600
>From: "Peter Siesjo" <peter.siesjo@neurokir.lu.se>
>Subject: Trues Blue
>
>Hello out there,
>Has anyone successfully stained something with True Blue. In our seek for a
>sensitive stain for cytokine detection in frozen sections we tried it. We
>were not able to get a  stain for more than a couple of seconds, then it
>disappeared despite different tricks recommended by the manufacturer. Or
>should it be renamed False Blue?
>Peter S
>
>Peter Siesjo, M.D., Ph.D.
>The Queen's Medical Center
>Center for the Study of Neurological Disease
>1356 Lusitana Street, U.T. 8th Floor
>Honolulu, HI 96813, USA
>Phone: (808) 537-7924
>Phone: (home) (808) 924 1653
>Fax:   (808) 537-7899
>Email:peter.siesjo@neurokir.lu.se
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 15:09:53 -0600
>From: Lynn Gardner <gardnerl@horus.ophth.uiowa.edu>
>Subject: Repeat messages
>
>To Whom It May Concern:
>
>The repeat messages are beginning again could someone please check into
>this. Thanks for your help!!
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 15:10:43 -0600
>From: "Kellar, Eric" <kellarec@MSX.UPMC.EDU>
>Subject: RE: Myeloperoxidase Staining
>
>Katie:
>
>Human myeloperoxidase (MPO) is an enzyme found in
>neutrophils/polymorphonuclear leucocytes (PMNs). MPO is a major
>component of the neutrophil host defense system, catalyzing the production
>of the microbiocidal oxidant, hypochlorous acid
>(HOCl), from chloride ions and hydrogen peroxide.
>
>Anti-MPO is a specific IPEX marker for neutrophils, and only labels the
>granules of neutrophil granulocytes and to a lesser extent, the granules of
>monocytes. It is released extracellularly (degranulation) after neutrophil
>activation in vitro or in vivo. Extracellular MPO may be an index of
>neutrophil activation in a variety of clinical conditions.
>
>Another technique is the enzyme histochemical method: Leder's Napthol AS-D
>chloroacetate esterase (CAE). The enzyme remains detectable after routine
>formalin fixation and paraffin embedding. The stain is (+) for mast cells
>and granulocytes, staining bright pink. It is (-) for other lymphoid cells.
>Mayer's hematoxylin is used as a counterstain and may suffice for your
>quantitation counts. It is more specific than anti-MPO but not as
sensitive.
>It really would be awesome to combine CAE or anti-MPO with a Giemsa
>technique, but the methanol would destroy the enzyme.
>
>A methyl green pyronin (MGP) may also be of interest to you. The MGP stains
>RNA red and DNA green to blue-green. The nucleoli will be red and the
>cytoplasm with many ribosomes as is found in plasma cells and transformed
>lymphocytes(especially of B cell type) and certain other cells will be
>bright red. Nuclei in general appear purple (or blue-green). The rest of
the
>backround is pale pink to colorless.
>
>
>Eric Kellar
>Histology/Immunohistochemistry
>University of Pittsburgh Medical Center
>
>
>
> ----------
> From:  Katie Bennett [SMTP:bresee@pilot.msu.edu]
> Sent:  Wednesday, December 30, 1998 11:58 AM
> To:  HistoNet Server
> Subject:  Re: Myeloperoxidase Staining
>
> To: Clif Chapman, Cathy Fragiskatos, and any other interested
>parties.
>
> What kind of antibody do you use against myeloperoxidase?  Does it
>stain
> neutrophils?
>
> I am trying the myeloperoxidase assay from the AFIP manual with
>little
> success.  Here's what I'm trying to do:
>
> I am trying to do a stain that will allow easy detection for
> quantitation of all the neutrophils in formalin-fixed,
>paraffin-embedded
> tissues so I can tell them apart from eosinophils and not rely
>solely on
> the morphology of the nucleus.  We have found that a May-Grunwald
>stain
> is great for the eos.  We probably could count the neutrophils in
>these
> May-Grunwald stained sections, but I would like to find something
>that
> makes those neutrophils really stand out.  I am not planning on
>doing
> both stains on one slide, but on serial sections.  However, a good
>IHC
> stain for neutrophils with a May-Grunwald counterstain would be
>awesome!
>
> Thanks in advance for any info you might have!
>
>
> Catherine "Katie" Bresee Bennett
> 218G Food Safety Toxicology Building
> Department of Pathology
> Michigan State University
> East Lansing, MI 48824
>
> ph:  (517) 432-4940
> fx:   (517) 353-9902
>
> bresee@pilot.msu.edu
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 15:45:54 -0600
>From: Cathy Mayton <histology@the-onramp.net>
>Subject:
>
>unsubscribe
>Cathy A. Mayton
>Wasatch Histo Consultants, Inc.
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 16:15:50 -0600
>From: rueggp@earthlink.net
>Subject: Re: Strange new artifact
>
>perhaps this is graphite from pencil labelling of cassetts or slides, or
>both.
>patsy ruegg
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 16:25:32 -0600
>From: larisonk@UONEURO.uoregon.edu
>Subject: Re: Trues Blue
>
>Peter S,
>
>True Blue has been used extensively by neurochemists to label retrogradely
>filled HRP-containing neurons.  In these experiments, ammonium molybdate is
>used to stabilize the signal.  One commonly cited reference for this
technique
>
>is Olucha, J Neuroscience Methods 13, 131-138 (1985).  It works quite well.
>Admittedly, I only tried the method once to develop an HRP-mediated
>immunohistochemical signal on frozen sections.  In this experiment, the
True
>Blue substrate was at least 4-fold more sensitive than the DAB substrate.
I
>next tried the technique on whole mount embryos, and that experiment was an
>abysmal failure.  So I haven't pursued it further.  My results comparing
True
>Blue and DAB were very preliminary.  For instance, I hadn't titered my
>antibody
>out far enough to get a true picture of True Blue's sensitivity.  Nor had I
>tried it with a variety of antibodies.  However, I've often wondered why it
>isn't used more extensively in the histochemical community.  I wondered
about
>the long-term stability of the signal.  After three months, the signal is
>still
>robust.  Does anyone know why this substrate isn't used more extensively?
>
>Karen D. Larison
>
>Date:          Wed, 30 Dec 1998 08:21:19 +0000
>From:          Peter =?iso-8859-1?Q?Siesj=F6?=
<peter.siesjo@neurokir.lu.se>
>Subject:       Trues Blue
>To:            histonet@pathology.swmed.edu
>
>Hello out there,
>Has anyone successfully stained something with True Blue. In our seek for a
>sensitive stain for cytokine detection in frozen sections we tried it. We
>were not able to get a  stain for more than a couple of seconds, then it
>disappeared despite different tricks recommended by the manufacturer. Or
>should it be renamed False Blue?
>Peter S
>
>Peter Siesjo, M.D., Ph.D.
>The Queen's Medical Center
>Center for the Study of Neurological Disease
>1356 Lusitana Street, U.T. 8th Floor
>Honolulu, HI 96813, USA
>Phone: (808) 537-7924
>Phone: (home) (808) 924 1653
>Fax:   (808) 537-7899
>Email:peter.siesjo@neurokir.lu.se
>
>
>
>
>
>
>
>----------------------------------------------------------------------
>
>Date: 30 Dec 1998 22:15:20 -0600
>From: poobear31@webtv.net (K M)
>Subject: Oh no! It's happening again!
>
>I'm again recieving messages anywhere from triplicate to pentuplate!  Is
>everyone else having this same problem again?
>
>
>
>Here are the messages received yesterday!
>