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Mark Sofferman
arme <@t> optonline.net
Tue Aug 19 08:18:51 CDT 2003
-----Original Message-----
From: HistoNet Server [mailto:histonet <@t> pathology.swmed.edu]
Sent: Tuesday, August 05, 2003 11:15 AM
To: HistoNet Server
Subject: Daily Digest
----------------------------------------------------------------------
Date: 4 Aug 2003 03:23:18 -0500
From: Chris van der Loos <c.m.vanderloos <@t> amc.uva.nl>
Subject: RE: more info again, End PO block
Dear Becky,
There is a new endogenous blocking solution from Dako called "Dual
endogenous enzyme block" (S2003). It will kill both endogenous peroxidase
and alkaline phosphatase activities in one 10 min incubation step prior to
your immunosteps. As this stuff is new my experience is only limited. So
far it seems very effective without damaging the tissue epitopes.
Furthermore, as Gayle Callis already wrote, methanol + 0.3% peroxide (20
min, RT) is very effective to block endogenous peroxidase activity in FFPE
sections. Because those sections have been trough alcohols several times
during embedding and dewaxing, the methanol has no additional damaging
effect. Be aware that methanol can be very damaging to many tissue epitopes
when immunostaining cryostat sections!
I have also tried to prolong the incubation time of a lesser effective
endogenous peroxidase blocking method as you wrote in your first mail, but
it didn't help.
Chris van der Loos
Dept. of Cardiovascular Pathology
Academical Medical Center
Amsterdam - The Netherlands
At 11:47 AM 8/1/2003 -0400, you wrote:
>I'm sorry about not giving enough information. I have had several
>responses asking for more details. So, I'll try again!
>
>I am performing IHC staining. I am using a DAKO autostainer with
>the LSAB detection system. I use DAB for my chromagen. My negative
>mouse and negative rabbit serum slides have quite specific staining
>in the granlocytic series of WBC's(No matter what pretreatment -
>I apply no pretreatment, Proteinase K, or use Microwave at pH 6 and
>10). Microwave pretreatment is the worst, but I get the staining
>most of the time. Polysegmented neutrophils seem to be the main
>culprit. Most of the times my Docs can read through this, but they
>are requesting my experimentation to find a "cure" for the situation,
>so they can honestly say, "no staining was seen in the negative
>control". One of the suggestions I have received is to do an Avidin/Biotin
>Block. I will give this a try, but if there are any other ideas out
>there: DON'T hold back! THANKS AGAIN!
----------------------------------------------------------------------
Date: 4 Aug 2003 05:05:13 -0500
From: "louise renton" <louise_renton <@t> hotmail.com>
Subject: Re: coated slides Elmer's
Dear all
I have heard a lot about Elmer's Glue, which I gather is some type of
household vinyl based adhesive. We don't get this specific brand, but
similar ones, used specifically for woodwork or crafts are available.
So...my question is this: How is it used? Neat, Diluted, in the
waterbath??? PLease advise as I have some deadful whole paw sections that I
am having difficuly "sticking" to the slide.
BTW, I have tried chrome alum, but get horrible background so my thanks go
to Gayle for suggesting placing them in NBF. I will try that too.
Best regards
Louise Renton
Bone Research Unit
MRC
Johannesburg
South Africa
Tel & fax +27 11 717 2298
"Time flies like an arrow, fruit flies like a banana"
- ----At 08:48 a.m. 01/08/2003 -0400, you wrote:
>Hi,
>I am having problems with tissue staying on my slides after drying
>overnight
>in 37C oven and then one more night in a 60C oven. The tissue is chicken
>leg knee joints. They are formalin fixed, EDTA decaled, and paraffin
>embedded samples. I think that coated slides may help. Does anyone have a
>simple protocol for making coated slides to help this tissue stick?
>Thanks in advance for the help.
>
>Loralee Gehan
>University of Rochester
> Loralle:
You can use a vinyl glue like Elmer#180#s, it works fine in cases
like you are describiyng. The adhesion properties are at least
the same that charged slides when you are treating samples
like cartilage and bone.
Good luck,Carlos.
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----------------------------------------------------------------------
Date: 4 Aug 2003 10:01:46 -0500
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: Caveat about Elmer's glue
Elmers glue has been used in the past, but some labs reported contamination
(bacterial, whatever) when using it - and if I remember correctly, there
have been publications/technical hints about this problem. That was enough
for our lab to abandon Elmers glue in lieu of in house prepared chrome
subbing solutions or (way back then) advent of poly l lysine or silane
coatings. We use Elmers glue for a short time, but disliked its white
opacity.
I was under the impression Elmers was a by product of the dairy industry,
since Bordens (of Elsie the Cow fame) manufactured this glue for school
children and household use. Personally, I would rather purchase
commercial(Surgipath) or make up chrome gelatin solution or some other
slide coating solution for histology purposes and leave Elmers to the
school children and households. When poly l lysine coating methods
arrived on the scene, we abandoned Elmers glue entirely or did the chrome
gelatin subbing for the very difficult, large decalcified sheep tibia, dog
tibia and femur section.
If you want to reduce background and are using 275 bloom gelatin, go to 100
bloom, larger molecule collagen often gives more H&E background staining
than smaller molecule (100 bloom) gelatin.
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
email: gcallis <@t> montana.edu
----------------------------------------------------------------------
Date: 4 Aug 2003 10:21:54 -0500
From: "DiCarlo, Margaret" <MDiCarlo <@t> KaleidaHealth.Org>
Subject: RE: coated slides Elmer's
Louise,
Fred told me about Titebond II which is an adhesive found in any local
hardware store. Just dilute it to make a 5% solution. I drop a few drops on
my slide using a disposable pipet and spread all over using kimwipe and air
dry it right before use and I have been having great results with it. There
is no background staining from using it. I hope you try this. Good luck.
Peggy DiCarlo HT(ASCP)
Orthopedics Bone Lab
Buffalo General Hospital
- -----Original Message-----
From: louise renton [mailto:louise_renton <@t> hotmail.com]
Sent: Monday, August 04, 2003 06:03
To: histonet <@t> pathology.swmed.edu
Subject: Re: coated slides Elmer's
Dear all
I have heard a lot about Elmer's Glue, which I gather is some type of
household vinyl based adhesive. We don't get this specific brand, but
similar ones, used specifically for woodwork or crafts are available.
So...my question is this: How is it used? Neat, Diluted, in the
waterbath??? PLease advise as I have some deadful whole paw sections that I
am having difficuly "sticking" to the slide.
BTW, I have tried chrome alum, but get horrible background so my thanks go
to Gayle for suggesting placing them in NBF. I will try that too.
Best regards
Louise Renton
Bone Research Unit
MRC
Johannesburg
South Africa
Tel & fax +27 11 717 2298
"Time flies like an arrow, fruit flies like a banana"
- ----At 08:48 a.m. 01/08/2003 -0400, you wrote:
>Hi,
>I am having problems with tissue staying on my slides after drying
>overnight
>in 37C oven and then one more night in a 60C oven. The tissue is chicken
>leg knee joints. They are formalin fixed, EDTA decaled, and paraffin
>embedded samples. I think that coated slides may help. Does anyone have a
>simple protocol for making coated slides to help this tissue stick?
>Thanks in advance for the help.
>
>Loralee Gehan
>University of Rochester
> Loralle:
You can use a vinyl glue like Elmer#180#s, it works fine in cases
like you are describiyng. The adhesion properties are at least
the same that charged slides when you are treating samples
like cartilage and bone.
Good luck,Carlos.
_________________________________________________________________
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----------------------------------------------------------------------
Date: 4 Aug 2003 10:47:21 -0500
From: Nick_Madary <@t> hgsi.com
Subject: Antibody for Anthrax Spores?
I am sure this has been discussed before but are there any commercially
available antibodies for anthrax spores?
******************* NOTE *******************
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contained in the following MIME Information.
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<br><font size=2 face="sans-serif">I am sure this has been discussed before
but are there any commercially available antibodies for anthrax
spores?</font>
- --=_alternative 0056194085256D78_=--
----------------------------------------------------------------------
Date: 4 Aug 2003 11:14:19 -0500
From: "Smith, Allen" <asmith <@t> mail.barry.edu>
Subject: RE: coated slides Elmer's
Since Elmer's is made by Borden's (a dairy company), it is probably casein.
Allen A. Smith, Ph.D.
Barry University
School of Graduate Medical Sciences
Podiatric Medicine and Surgery
Miami Shores, Florida 33161-6695
- -----Original Message-----
From: louise renton [mailto:louise_renton <@t> hotmail.com]
Sent: Monday, August 04, 2003 6:03 AM
To: histonet <@t> pathology.swmed.edu
Subject: Re: coated slides Elmer's
Dear all
I have heard a lot about Elmer's Glue, which I gather is some type of
household vinyl based adhesive. We don't get this specific brand, but
similar ones, used specifically for woodwork or crafts are available.
So...my question is this: How is it used? Neat, Diluted, in the
waterbath??? PLease advise as I have some deadful whole paw sections that I
am having difficuly "sticking" to the slide.
BTW, I have tried chrome alum, but get horrible background so my thanks go
to Gayle for suggesting placing them in NBF. I will try that too.
Best regards
Louise Renton
Bone Research Unit
MRC
Johannesburg
South Africa
Tel & fax +27 11 717 2298
"Time flies like an arrow, fruit flies like a banana"
- ----At 08:48 a.m. 01/08/2003 -0400, you wrote:
>Hi,
>I am having problems with tissue staying on my slides after drying
>overnight
>in 37C oven and then one more night in a 60C oven. The tissue is chicken
>leg knee joints. They are formalin fixed, EDTA decaled, and paraffin
>embedded samples. I think that coated slides may help. Does anyone have a
>simple protocol for making coated slides to help this tissue stick?
>Thanks in advance for the help.
>
>Loralee Gehan
>University of Rochester
> Loralle:
You can use a vinyl glue like Elmer#180#s, it works fine in cases
like you are describiyng. The adhesion properties are at least
the same that charged slides when you are treating samples
like cartilage and bone.
Good luck,Carlos.
_________________________________________________________________
Unsatisfied with being single? Try MSN Personals
http://www.msn.co.za/personals/
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----------------------------------------------------------------------
Date: 4 Aug 2003 11:57:39 -0500
From: "Cathrine Dreanno" <Catherine.Dreanno <@t> newcastle.ac.uk>
Subject: RDO-gold and ISH
Hi,
I would like to decalcify my samples and do some in situ hybridization. I
read that RDO is not suitable for ISH (Alers et al., 1999, 47; 703-709,
JHC).
They have a new product called "RDO-gold". They say the "nucleic acid are
altered". Does anyone have any experience of this product ?
Thank you for advices ,
Catherine
----------------------------------------------------------------------
Date: 4 Aug 2003 12:20:35 -0500
From: Margaret_McKinney <@t> brown.edu
Subject: flash freezing, brain, and isopentane
I have a quick question regarding flash freezing of rat brain in freezing
isopentane. Does anyone have an opinion on flash-freezing by simply dropping
the brain into the isopentane versus placing it in a container with OCT or
similar media and then placing this into the isopentane? I have been running
into problems with both micro and macro cracking of the tissue.
Thanks,
Margaret McKinney.
Margaret McKinney
Brown University Medical School
97 Waterman St.
Providence, RI 02912
----------------------------------------------------------------------
Date: 4 Aug 2003 12:48:52 -0500
From: Judi Ford <jlf <@t> jax.org>
Subject: Gallyas Stain on Brain
Hi,
I've just finished doing the Gallyas stain for tangles and have found that
no
matter how gentle I am with the slides tissue falls off the slide during the
stain. I thought about using + slides but heard that brain tissue and plus
slides don't work due to +/- charges working against each other. Any
suggestions? Also, what could I use as a control for tangles? I'm working
with mouse tissue.
Thanks for your help.
Judi Ford
Histotechnologist
Jackson Laboratory, ME
----------------------------------------------------------------------
Date: 4 Aug 2003 12:49:14 -0500
From: "Dawson, Glen" <GDawson <@t> Milw.Dynacare.com>
Subject: MIB-1 (Ki-67) VENDOR
All,
I am looking for the MIB-1 (Ki-67) antibody that used to be sold by
Immunotech (Cat. #IM0505). Unless I'm mistaken, Immunotech is no longer
around but this antibody is/was the best. Any leads on who may actually
have this particular antibody or who may have bought out immunotech would be
much appreciated.
Thanx in Advance,
Glen A. Dawson BS, HT & QIHC (ASCP)
Lead IHC Technologist
Milwaukee, WI
----------------------------------------------------------------------
Date: 4 Aug 2003 12:49:41 -0500
From: Nick_Madary <@t> hgsi.com
Subject: Re: flash freezing, brain, and isopentane
Snap freeze the whole brain as it is almost its own "oct" type matrix. You
wouls still want to use the oct as a method for sticking the brain onto
the chuck or block holder.
Margaret_McKinney <@t> brown.edu
08/04/03 12:16 PM
To: histonet <@t> pathology.swmed.edu
cc:
Subject: flash freezing, brain, and isopentane
I have a quick question regarding flash freezing of rat brain in freezing
isopentane. Does anyone have an opinion on flash-freezing by simply
dropping the brain into the isopentane versus placing it in a container
with OCT or similar media and then placing this into the isopentane? I
have been running into problems with both micro and macro cracking of the
tissue.
Thanks,
Margaret McKinney.
Margaret McKinney
Brown University Medical School
97 Waterman St.
Providence, RI 02912
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contained in the following MIME Information.
********************************************
- ------------------ MIME Information follows ------------------
This is a multipart message in MIME format.
- --=_alternative 005E974405256D78_=
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<br><font size=2 face="sans-serif">Snap freeze the whole brain as it is
almost
its own "oct" type matrix. You wouls still want to use the oct as
a
method for sticking the brain onto the chuck or block holder.</font>
<br>
<br>
<br>
<table width=100%>
<tr valign=top>
<td>
<td><font size=1 face="sans-serif"><b>Margaret_McKinney <@t> brown.edu</b></font>
<p><font size=1 face="sans-serif">08/04/03 12:16 PM</font>
<br>
<td><font size=1 face="Arial"> </font>
<br><font size=1 face="sans-serif"> To:
histonet <@t> pathology.swmed.edu</font>
<br><font size=1 face="sans-serif"> cc:
</font>
<br><font size=1 face="sans-serif"> Subject:
flash freezing, brain, and isopentane</font></table>
<br>
<br>
<br><font size=2><tt>I have a quick question regarding flash freezing of rat
brain in freezing isopentane. Does anyone have an opinion on flash-freezing
by
simply dropping the brain into the isopentane versus placing it in a
container
with OCT or similar media and then placing this into the isopentane? I have
been running into problems with both micro and macro cracking of the
tissue.<br>
<br>
Thanks,<br>
Margaret McKinney.<br>
<br>
Margaret McKinney<br>
Brown University Medical School<br>
97 Waterman St.<br>
Providence, RI 02912<br>
<br>
<br>
</tt></font>
<br>
<br>
- --=_alternative 005E974405256D78_=--
----------------------------------------------------------------------
Date: 4 Aug 2003 13:58:50 -0500
From: "Jocelyn Torcolini" <jmt175 <@t> psu.edu>
Subject: RE:
unsubscribe
----------------------------------------------------------------------
Date: 4 Aug 2003 14:00:12 -0500
From: "Bryan Hewlett" <bhewlett <@t> cogeco.ca>
Subject: Re: MIB-1 (Ki-67) VENDOR
Glen,
This clone is available from DakoCytomation.
Bryan
- ----- Original Message -----
From: "Dawson, Glen" <GDawson <@t> Milw.Dynacare.com>
To: <histonet <@t> pathology.swmed.edu>
Sent: Monday, August 04, 2003 1:37 PM
Subject: MIB-1 (Ki-67) VENDOR
>
> All,
>
> I am looking for the MIB-1 (Ki-67) antibody that used to be sold by
> Immunotech (Cat. #IM0505). Unless I'm mistaken, Immunotech is no longer
> around but this antibody is/was the best. Any leads on who may actually
> have this particular antibody or who may have bought out immunotech would
be
> much appreciated.
>
> Thanx in Advance,
>
> Glen A. Dawson BS, HT & QIHC (ASCP)
> Lead IHC Technologist
> Milwaukee, WI
>
>
----------------------------------------------------------------------
Date: 4 Aug 2003 14:01:01 -0500
From: "Yaskovich, Ruth A (NIH/NIDCR)" <ryaskovich <@t> dir.nidcr.nih.gov>
Subject: RE: Gallyas Stain on Brain
I cut mouse brain all the time on plus slides from statlabs with no problem
with tissue falling off.
Ruth Yaskovich
- -----Original Message-----
From: Judi Ford [mailto:jlf <@t> jax.org]
Sent: Monday, August 04, 2003 1:34 PM
To: histonet <@t> pathology.swmed.edu
Subject: Gallyas Stain on Brain
Hi,
I've just finished doing the Gallyas stain for tangles and have found that
no matter how gentle I am with the slides tissue falls off the slide during
the stain. I thought about using + slides but heard that brain tissue and
plus slides don't work due to +/- charges working against each other. Any
suggestions? Also, what could I use as a control for tangles? I'm working
with mouse tissue.
Thanks for your help.
Judi Ford
Histotechnologist
Jackson Laboratory, ME
----------------------------------------------------------------------
Date: 4 Aug 2003 14:01:43 -0500
From: "Wright, Barbara (DNAX)" <barbara.wright2 <@t> dnax.org>
Subject: RE: MIB-1 (Ki-67) VENDOR
Glen,
I belive you can get Immunotech products through Becton Dickinson.
DAKO's Ki-67 MIB-1 antibody works great too.
Hope that helps
Barb Wright
- -----Original Message-----
From: Dawson, Glen [mailto:GDawson <@t> Milw.Dynacare.com]
Sent: Monday, August 04, 2003 10:38 AM
To: histonet <@t> pathology.swmed.edu
Subject: MIB-1 (Ki-67) VENDOR
All,
I am looking for the MIB-1 (Ki-67) antibody that used to be sold by
Immunotech (Cat. #IM0505). Unless I'm mistaken, Immunotech is no longer
around but this antibody is/was the best. Any leads on who may actually
have this particular antibody or who may have bought out immunotech would be
much appreciated.
Thanx in Advance,
Glen A. Dawson BS, HT & QIHC (ASCP)
Lead IHC Technologist
Milwaukee, WI
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